Immunomodulatory compositions and methods of use thereof

ABSTRACT

The present disclosure provides immunomodulatory compositions comprising live Caulobacter crescentus (CC). Immunomodulatory compositions of the present disclosure are useful for modulating an immune response in an individual. The present disclosure thus provides methods of modulating an immune response in an individual, involving administering an immunomodulatory composition comprising live CC to the individual.

INTRODUCTION

Caulobacter crescentus is non-pathogenic, harmless, aquatic,gram-negative bacterium that grows at ˜23° C. in many soil andfreshwater environments Caulobacter has been studied for nearly 50years. The main laboratory strain (C. crescentus CB15) is wellcharacterized genetically and biochemically, and the genome of C.crescentus has been sequenced. Caulobacters are readily grown usingstandard laboratory equipment. They can also be easily grown incommercial fermenters to at least 30 optical density units (ODs) inanimal protein free, defined minimal media.

There is a need in the art for immunomodulatory compositions that can beused in the prophylaxis and/or treatment of disorders such asinflammation, undesirable inflammatory activity, exacerbated immuneresponses, aberrant immune responses, immune dysregulation andautoimmune diseases.

SUMMARY

The present disclosure provides immunomodulatory compositions comprisingCaulobacter crescentus (CC). Immunomodulatory compositions of thepresent disclosure are useful for modulating an immune response in anindividual. The present disclosure provides methods of modulating animmune response in an individual, involving administering animmunomodulatory composition comprising CC to the individual.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 demonstrates the reduction in concanavalin A (ConA) inducedpro-inflammatory cytokines (IFN-γ, TNF-α, IL-6 and IL-17A) production insplenocytes isolated from CC and PBS treated mice by intranasal and oralroutes twice weekly. Data are expressed in pg/ml and shown asaverage±standard deviation (SD) of triplicates.

FIG. 2 demonstrates the reduction in pokeweed mitogen (PWM) inducedpro-inflammatory cytokines (IFN-γ, TNF-α, IL-6 and IL-17A) production insplenocytes isolated from CC and phosphate buffered saline (PBS) treatedmice by intranasal and oral routes twice weekly. Data are expressed inpg/ml and shown as average+SD of triplicates.

FIG. 3A-3C illustrate the induction of IL-10 in splenocytes isolatedfrom CC and PBS treated mice by intranasal and oral routes twice weeklyand stimulated ex vivo for 24 h with medium (A), PWM (B) and ConA (C).Data are expressed in pg/ml and shown as average+SD of triplicates.

FIG. 4 illustrates the induction profile of IL-10 in splenocytesisolated from CC and PBS treated mice orally twice a week. Cells werestimulated in vitro with LPS for 24 or 72 hrs.

FIG. 5 shows the reduction profile of cytokines (IFN-γ, IL-6 and IL-17A)in mesenteric lymph nodes isolated from CC and PBS administered miceorally twice a week. Cells were stimulated in vitro withlipopolysaccharide (LPS) for 24 hrs. Data are shown as average+SD.

FIG. 6 illustrates the enhancement of IL-10 producing CD4+ and CD8+ Tcells in splenocytes isolated from CC and PBS fed mice twice weekly.Percent of cells positive for CD3 and CD4 or CD8, expressingintracellular IL-10 are presented.

FIG. 7 demonstrates the modulation in ConA induced pro-inflammatorycytokines (IFN-γ, TNF-α and IL-6) production in splenocytes isolatedfrom CC and PBS treated mice by subcutaneous route once weekly. Data areexpressed in pg/ml and shown as average+SD of triplicates.

FIG. 8 demonstrates the induction of regulatory lymphocytes (CD8 Tcells, NKT cells and NK cells) in mice treated with CC or PBS orallyonce weekly. The data represent CD3⁺CD8⁺, CD3+CD49b+ and CD3-CD49b⁺populations expressing CD25, FoxP3 or PD-1.

FIG. 9 demonstrates the induction of regulatory lymphocytes (CD4 and CD8T cells) in mice treated with CC or PBS subcutaneously once weekly. Thedata represent CD3⁺CD4⁺ and CD3⁺CD8⁺ populations expressing CD25 andFoxP3.

FIG. 10 shows the modulation of IL-113, IL-6 and IL-10 in sera samplesfrom CC and PBS treated mice orally post 2 hours (hrs) in vivo challengewith LPS at 7 mg/Kg. The LPS challenged mice has dramatic increase ininflammatory cytokines in sera, which were reduced in mice treated withCC. In contrast CC treatment led to an increase in IL-10 levels in LPSchallenged mice.

FIG. 11 demonstrates the modulation of TNF-α, IL-6, IL-1β, and IL-17A inliver and lung homogenate samples from CC and PBS treated mice orallypost 2 hrs in vivo challenge with LPS at 7 mg/kg. CC used was preparedin two formats: CC-1 (CC grown in liquid peptone yeast extract (PYE)medium and stored at room temperature in saline), and CC-2 (CC grown inliquid PYE medium and stored at 4° C. in refrigerator). Data representaverage+SD of triplicate wells.

FIG. 12 illustrates the biochemical parameters (GLU: glucose; GLOB:Globulin; ALT: Alanine aminotransferases; TP: total phosphates; TBIL:total bilirubin) in the sera samples from CC and PBS treated mice orallypost 2 hrs in vivo challenge with LPS at 7 mg/Kg. CC used was preparedin two formats: CC-1 (CC grown in liquid PYE medium and stored at roomtemperature in saline), and CC-2 (CC grown in liquid PYE medium andstored at 4° C. in refrigerator).

FIG. 13 shows hematoxylin and eosin (H& E) staining of the liversections isolated from CC and PBS treated mice orally post 2 hrs in vivochallenge with LPS at 7 mg/Kg. CC used was prepared in two formats: CC-1(CC grown in liquid PYE medium and stored at room temperature insaline), and CC-2 (CC grown in liquid PYE medium and stored at 4° C. inrefrigerator).

FIG. 14 demonstrates the modulation of cytokines (IFN-γ, IL-6, IL-17Aand IL-22) in Aldra (Imiquimod) induced psoriasis mouse model upon twoweek (5 days/wk) oral treatment with CC and PBS. Splenocytes wereharvested from mice and cultured ex vivo for 4 days in medium, followedby collecting supernatant and testing cytokines.

FIG. 15 demonstrates the modulation of cytokines (IFN-γ, TNF-α, IL-1β,IL-8, MIP-1α) in DSS (Dextran sulfate sodium) induced IBD (inflammatorybowel disease) mouse model. Data are expressed in pg/ml and shown asaverage±SD of triplicates.

FIG. 16 demonstrates the suppression of autoantigen specific T cellproliferation and antibody (IgG and IgG2a) responses in an experimentalautoimmune encephalomyelitis (EAE) mouse model upon oral treatment withCC.

FIG. 17 demonstrates the suppression of allergen specific IgE antibodyresponses in an ovalbumin-induced allergic airway inflammation modelupon oral treatment with CC.

FIG. 18 demonstrates reduction of allergen specific cytokines (IL-4 andIL-6) in spleen in an ovalbumin-induced allergic airway inflammationmodel upon oral treatment with CC. Data are expressed in pg/ml and shownas average±SD of triplicates.

FIG. 19 illustrates reduction of allergen specific cytokines (IL-4 andIL-6) in spleen in an ovalbumin-induced allergic airway inflammationmodel upon oral treatment with CC+dexamethasone (DEX), DEX alone or notreatment control. Data are expressed in pg/ml and shown as average±SDof triplicates.

FIG. 20 demonstrates the effect of oral treatment with CC onpro-inflammatory cytokines (IFN-γ, TNF-α, IL-6 and IL-1β) in liver ofmice on high fat diet, compared to untreated high-fat diet fed mice.Data were obtained at ˜180 days after initiation of high-fat diet andare shown as mean±SD of 5 mice. Data are expressed in pg/ml and shown asaverage±SD of triplicates.

FIG. 21 shows the effect of oral treatment with CC on pro-inflammatorycytokines (IFN-γ, TNF-α and IL-6) in spleen of mice on high fat diet,compared to untreated high-fat diet fed mice. Data were obtained at ˜180days after initiation of high-fat diet and are shown as mean±SD of 5mice. Data are expressed in pg/ml.

FIG. 22 illustrates the biochemical parameters (CHOL: cholesterol, TRIG:triglycerides, URIC: uric acid, CK: creatine kinase) in the sera samplesfrom CC treated mice on high fat diet, and compared to untreated highfat diet fed mice. Data were obtained at ˜180 days after initiation ofhigh-fat diet and are shown as mean of 5 mice.

FIG. 23 illustrates the effect of CC on glucose tolerance in mice onhigh fat diet, compared to untreated high-fat diet fed mice. Data wereobtained at ˜180 days after initiation of high-fat diet and are shown asmean±SD of 5 mice.

FIG. 24 illustrates the biochemical parameters (PHOS: phosphate; TBIL:total bilirubin) in the sera samples from CC+cyclophosphamide andcyclophosphamide treated mice bearing EL-4 subcutaneous tumor, andcompared to normal mice.

FIG. 25 illustrates the biochemical parameters (PHOS: phosphate; UREA;TBIL: total bilirubin; AST: aspartate aminotransferase) in the serasamples from CC+cisplatin and cisplatin treated mice with B16 metastaticcancer, and compared to normal mice.

FIG. 26 illustrates the biochemical parameters (ALT: alanineaminotransferases; AST: aspartate aminotransferase) in the sera samplesfrom CC+anti-PD-1 monoclonal antibody and anti-PD-1 monoclonal antibodytreated mice with B16 tumor, and compared to normal mice.

FIG. 27 demonstrates the modulation of IFN-γ, TNF-α, IL-6, IL-1β,MIP-1α, IL-8 and IL-10 in liver homogenate samples from LPS^(−ve) CC andPBS treated mice orally post 2 and 24 hrs in vivo challenge with LPS at25 mg/Kg, i.p. Data are expressed in pg/ml and represent average±SD oftriplicate wells.

FIG. 28 demonstrates that Caulobacter vibroides (CV) can lead to downregulation of inflammatory cytokines (IFN-γ and TNF-α) induced by aprobiotic (Lactobacillus rhamnosus, LB) or a pathogenic bacterium(Lysteria monocytogenes, LM) from human PBMCs in cell cultures. Data areexpressed in pg/ml and represent average+SD of triplicate wells.

FIG. 29 shows that CC induces IL-10 production from human myeloiddendritic cells (DCs) cultured ex vivo. Data are expressed in pg/ml andrepresent average±SD of triplicate wells.

FIG. 30 demonstrates that CC can be used to differentiate/expand myeloidcells from pluripotent stem cells present in human peripheral blood. Thedata represent CD34⁺CD45⁻, CD34⁺CD45⁻CD11c⁺ and CD34⁺CD45⁻CD11b⁺populations as determined by flow cytometry.

DEFINITIONS

The terms “individual,” “host,” “subject,” and “patient” are usedinterchangeably herein, and refer to mammals, including, but not limitedto, humans, non-human primates (e.g. simians), non-human mammals (e.g.,mammalian livestock animals (e.g., bovine, porcine, caprine, and ovineanimals)), and mammalian pets (e.g., cats, dogs); fish; and birds (e.g.,chicken).

A “biological sample” encompasses a variety of sample types obtainedfrom an individual. The definition encompasses blood, serum, plasma, andother liquid samples of biological origin; solid tissue samples such asa biopsy specimen or tissue cultures or cells derived therefrom and theprogeny thereof. The definition also includes samples that have beenmanipulated in any way after their procurement, such as by treatmentwith reagents; washed; or enrichment for certain cell populations, suchas epithelial cells. The term “biological sample” encompasses a clinicalsample, and also includes cells in culture, cell supernatants, organs,tissue samples, lung biopsy samples, lung epithelial cells,gastrointestinal epithelial cells, gastrointestinal tract tissuesamples, bronchoalveolar lavage (BAL) fluid samples, nasal lavage fluidsamples, blood, plasma, serum, cerebrospinal fluid, fecal samples, andthe like.

An “immunomodulator” or “immunomodulatory agent” is any agent which doesone or more of: restores depressed immune function, down-regulates anabnormal immune function, regulates abnormal/excessive immune function,enhances normal immune function, and provide desired immune response.Immune function includes one or more of: humoral (antibody-mediated)immunity, cellular immunity, and innate immunity. An “immunomodulator”includes agents acting directly on the cells involved in the expressionof immune response, or on cellular or molecular mechanisms, which, inturn, act to modify the function of cells involved in immune response.Regulation of immune function may result from the action of animmunomodulatory agent to abrogate activating mechanisms derived bypositive-feedback influences endogenous or exogenous to the immunesystem. Regulation of immune function may result from the action of animmunomodulatory agent to abrogate suppressing mechanisms derived bynegative-feedback influences endogenous or exogenous to the immunesystem. Thus, immunomodulators can have diverse mechanisms of action.

The terms “modulate” and “modulation” refer to increasing, reducing, orbalancing the number and/or activity of immune cells, cytokines,chemokines, antibodies, soluble factors, surface molecules,intracellular molecules, effector functions, regulatory functions etc.

The terms “autoimmune disease” and “autoimmune disorder” are usedinterchangeably herein to refer to diseases characterized by an immuneresponse to a self antigen, i.e., an immune response to substances andtissues normally present in the body.

The term “inflammatory disease” refers to a disease caused by, resultingfrom, or resulting in inflammation. The term “inflammatory disease” mayalso refer to a dysregulated inflammatory reaction that causes anexaggerated response by various innate and adaptive immune cells leadingto abnormal tissue damage and/or cell death.

An “adjuvant” is any agent which is capable of potentiating an immuneresponse and are, therefore, one class of immunopotentiators (Stites andTen, Basic and Clinical Immunology, 7^(th) Ed., Appleton and LangeNorwalk Conn. pp. 797, 1991). Adjuvants are used to increase the immuneresponses in vaccination in order to enhance the humoral and/or cellmediated immune responses.

A “cytokine” means any secreted polypeptide that affects the functionsof other cells, and is a molecule, which modulates interactions betweencells in the immune or inflammatory response. A cytokine includes, butis not limited to monokines, chemokines, and lymphokines, regardless ofwhich cells produce them.

The terms “antibodies” and “immunoglobulin” include antibodies orimmunoglobulins of any isotype, fragments of antibodies which retainspecific binding to antigen, including, but not limited to, Fab, Fv,scFv, and Fd fragments, chimeric antibodies, humanized antibodies,single-chain antibodies, bi-specific antibodies, and fusion proteinscomprising an antigen-binding portion of an antibody and a non-antibodyprotein. Also encompassed by the term are Fab′, Fv, F(ab′)₂, and orother antibody fragments that retain specific binding to antigen, andmonoclonal antibodies. An antibody may be monovalent or bivalent.

A “therapeutically effective amount” or “efficacious amount” means theamount of a compound or agent that, when administered to a mammal orother subject for treating a disease, is sufficient to effect suchtreatment for the disease. The “therapeutically effective amount” willvary depending on the compound or agent, the disease and its severityand the age, weight, general health status, sex, etc., of the subject tobe treated. In some cases, an “effective amount” of an agent is anamount that: 1) restores the immune function to normal levels; 2)modulates immune function to normal levels; or 3) reduces immunefunction to below a pathological level.

The terms “treatment”, “treating” and the like are used herein togenerally mean obtaining a desired pharmacologic and/or physiologiceffect. The effect may be prophylactic in terms of completely orpartially preventing a disease or symptom thereof and/or may betherapeutic in terms of a partial or complete cure for a disease and/oradverse effect attributable to the disease. “Treatment” as used hereincovers any treatment of a disease or symptom in a mammal, and includes:(a) preventing the disease or symptom from occurring in a subject whichmay be predisposed to acquiring the disease or symptom but has not yetbeen diagnosed as having it; (b) inhibiting the disease or symptom,i.e., arresting its development; or (c) relieving the disease, i.e.,causing regression of the disease. The therapeutic agent may beadministered before, during or after the onset of disease or injury. Thetreatment of ongoing disease, where the treatment stabilizes or reducesthe undesirable clinical symptoms of the patient, is of particularinterest. Such treatment is desirably performed prior to complete lossof function in the affected tissues. The subject therapy will desirablybe administered during the symptomatic stage of the disease, and in somecases after the symptomatic stage of the disease.

A “pharmaceutically acceptable carrier or excipient” means a non-toxicsolid, semi-solid, or liquid filler, diluent, encapsulating material orformulation auxiliary of any type. One skilled in the art of preparingformulations can readily select the proper form and mode ofadministration depending upon the particular characteristics of the gentselected, the disease state to be treated, the stage of the disease, andother relevant circumstances.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. Thus, for example, reference to “alive Caulobacter crescentus” includes a plurality of such bacteria andreference to “the cytokine” includes references to one or more cytokinesand equivalents thereof known to those skilled in the art, and so forth.It is further noted that the claims may be drafted to exclude anyoptional element. As such, this statement is intended to serve asantecedent basis for use of such exclusive terminology as “solely,”“only” and the like in connection with the recitation of claim elements,or use of a “negative” limitation.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination. All combinations of the embodimentspertaining to the invention are specifically embraced by the presentinvention and are disclosed herein just as if each and every combinationwas individually and explicitly disclosed. In addition, allsub-combinations of the various embodiments and elements thereof arealso specifically embraced by the present invention and are disclosedherein just as if each and every such sub-combination was individuallyand explicitly disclosed herein.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

DETAILED DESCRIPTION

The present disclosure relates to Caulobacter crescentus (CC) and itsuse as an immunomodulatory biotherapeutic agent. CC has been shown tohave immunomodulatory effects by modulating innate and adaptive immuneresponses. The present disclosure provides immunomodulatory compositionscomprising live Caulobacter crescentus (CC). Immunomodulatorycompositions of the present disclosure are useful for modulating animmune response in an individual. Immunomodulatory compositions of thepresent disclosure are useful for reducing an undesired immune responsein an individual. Immunomodulatory compositions of the presentdisclosure are useful for reducing inflammation in an individual. Thepresent disclosure provides methods of modulating an immune response inan individual, involving administering an immunomodulatory compositioncomprising CC to the individual. The present disclosure provides methodsof reducing an undesired immune response in an individual, involvingadministering an immunomodulatory composition comprising live CC to theindividual. The present disclosure provides methods of reducinginflammation in an individual, involving administering animmunomodulatory composition comprising live CC to the individual.

Immunomodulatory Compositions

The present disclosure provides immunomodulatory compositions comprisingCaulobacter crescentus (CC). CC in an immunomodulatory composition ofthe present disclosure are viable. CC in an immunomodulatory compositioncan be non-denatured, mutated, attenuated and/or genetically modified.An immunomodulatory composition of the present disclosure can comprise acocktail of one or more different strains of Caulobacter crescentusbacteria.

CC-containing immunomodulatory compositions include the CC by itselfwith a pharmaceutically acceptable carrier or excipients forimmunomodulatory activity, including “adjuvanting” in which CCadministration to a subject may be wholly independent of, and/orseparated temporally and/or spatially from, administration to thesubject of one or more antigens against which modulation or regulationof an immune response (e.g., an antigen specific response) in thesubject is desired.

An immunomodulatory composition of the present disclosure can modulate(e.g., reduce) an immune response in an individual. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number of Bcells in an individual. For example, in some cases, an effective amountof an immunomodulatory composition of the present disclosure is anamount that is effective, when administered in a single dose or inmultiple doses, to modulate (e.g., reduce) the number of B cells in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, or more than 10-fold, compared to the numberof B cells in the individual in the absence of treatment with theimmunomodulatory composition. In some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) the number of antigen-specific B cells in anindividual. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number of antigen-specific B cells in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number of antigen-specific B cells in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number of autoantigen-specific B cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number of autoantigen-specific B cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number of allergen-specific B cells in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number of allergen-specific B cells inthe individual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce)activity of B cells in an individual. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) activation of Bcells in an individual by at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold,at least 100-fold, or more than 100-fold, compared to the activationlevel of B cells in the individual in the absence of treatment with theimmunomodulatory composition. For example, in some cases, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to reduce the activation of B cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the activation level of B cells in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) theamount of antibody specific to a given antigen in the individual. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate(e.g., reduce) the amount of antibody specific to a given antigen in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, or more than 10-fold, compared to the amountof antibody specific to the antigen in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the amount of antibodyspecific to a given antigen in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of antibody specific to the antigen in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the amount of autoantigen-specific antibody in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the amount of autoantigen-specific antibodyin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the amount of allergen-specific antibody in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the amount of allergen-specific antibody inthe individual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce)production of one or more cytokines in the individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce)production of one or more cytokines in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, or morethan 10-fold, compared to the amount of the cytokine in the individualin the absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce theproduction of one or more cytokines in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the production of one or more cytokines in the individual inthe absence of treatment with the immunomodulatory composition. In othercases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to modulate (e.g., reduce)production of GM-CSF in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, or more than 10-fold,compared to the amount of GM-CSF in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the production of GM-CSFin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the amount of GM-CSF inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) production of IL-22 in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, or more than 10-fold, compared to the amount of IL-22 in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of IL-22 in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of IL-22 in the individual in the absence oftreatment with the immunomodulatory composition. In other, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to modulate (e.g., reduce) production of interferon(IFN)-α and/or IFN-β and/or IFN-γ in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, or morethan 10-fold, compared to the amount of IFN-α or IFN-β or IFN-γ in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of interferon (IFN)-α and/or IFN-β and/or IFN-γin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the amount ofinterferon (IFN)-α and/or IFN-β and/or IFN-γ in the individual in theabsence of treatment with the immunomodulatory composition. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to reduce the production of IFN-γin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the amount of IFN-γ inthe individual in the absence of treatment with the immunomodulatorycomposition.

As another example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) production of one or more of IL-1β, IL-17A,IL-2, IL-10, IL-6 and TNF-α in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, or morethan 10-fold, compared to the amount of IL-1β, IL-17A, IL-2, IL-10,IL-6, or TNF-α in the individual in the absence of treatment with theimmunomodulatory composition. In some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to increase the level of IL-10 in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, or morethan 10-fold, compared to the amount of IL-10 in the individual in theabsence of treatment with the immunomodulatory composition. In somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to increase the number ofIL-10-producing CD4⁺ and/or CD8⁺ T cells in an individual by at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45%, at least 50%, at least 75%, at least100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold,or more than 10-fold, compared to the number of IL-10-producing CD4⁺and/or CD8⁺ T cells in the individual in the absence of treatment withthe immunomodulatory composition. As another example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to reduce the production of one or more ofIL-β, IL-17A, IL-2, IL-6 and TNF-α in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of one or more of IL-1β, IL-17A, IL-2, IL-6 andTNF-α in the individual in the absence of treatment with theimmunomodulatory composition. For example, in some cases, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to reduce the production of IL-17 in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the amount of IL-17 in the individual in theabsence of treatment with the immunomodulatory composition. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to reduce the production of IL-2in an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the amount of IL-2 theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of TNF-α in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of TNF-α in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the production of IL-6 inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the amount of IL-6 in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of IL-1β in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of IL-1β in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to increase the level of TGF-β in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, or more than 10-fold, compared to the amountof TGF-β in the individual in the absence of treatment with theimmunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., increase,reduce or balance) production of one or more cytokines, chemokines orlymphotoxins such as but not limited to GM-CSF, IL-2, IL-22,Interferons, IL-1β, TGF-β, IL-17A, IL-2, IL-10, IL-6, IL-5, IL-13,TNF-α, IL-9, IL-28, KC/IL-8, MIP-1α, LTa4 etc. in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, or more than 10-fold, compared to the amount of cytokines,chemokines or lymphotoxins such as but not limited to GM-CSF, IL-2,IL-22, Interferons, IL-1β, TGF-β, IL-17A, IL-2, IL-10, IL-6, IL-5,IL-13, TNF-α, IL-9, IL-28, KC/IL-8, MIP-1α, LTα4 etc. in the individualin the absence of treatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) a Th1response in an individual. For example, in some cases, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to modulate (e.g., reduce) a Th1 response in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the level of the Th1 response in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce a Th1 response in an individual by at least 10%, at least 15%,at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, or more than 75%, compared to theTh1 response in the individual in the absence of treatment with theimmunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD4⁺ T cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD4+ T cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of CD4⁺ T cells in the individual in the absenceof treatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific CD4⁺ T cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific CD4⁺ T cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific CD4⁺ T cellsin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of CD4+ T cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of CD4⁺ T cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific CD4⁺ T cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific CD4⁺ T cells in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the number and/or activityof autoantigen-specific CD4⁺ T cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the number and/or activity of autoantigen-specific CD4+ Tcells in the individual in the absence of treatment with theimmunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD8⁺ T cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD8⁺ T cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of CD8⁺ T cells in the individual in the absenceof treatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific CD8⁺ T cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific CD8⁺ T cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific CD8⁺ T cellsin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of CD8⁺ T in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of CD8⁺ T in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific CD8⁺ T in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific CD8⁺ T in the individual in the absence of treatmentwith the immunomodulatory composition. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to reduce the number and/or activity ofautoantigen-specific CD8⁺ T cells in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the number and/or activity of autoantigen-specific CD8⁺ Tcells in the individual in the absence of treatment with theimmunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of cytolytic T cells in an individual. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate(e.g., reduce) the number and/or activity of cytolytic T cells in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number and/or activity of cytolytic Tcells in the individual in the absence of treatment with theimmunomodulatory composition. In some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) the number and/or activity ofantigen-specific cytolytic T cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific cytolytic T cells in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number and/or activity ofantigen-specific cytolytic T cells in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the number and/or activityof cytolytic T cells in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, or more than 75%, compared to thenumber and/or activity of cytolytic T cells in the individual in theabsence of treatment with the immunomodulatory composition. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to reduce the number and/oractivity of antigen-specific cytolytic T cells in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity ofantigen-specific cytolytic T cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate the number and/oractivity of one or more of natural killer (NK) cells, NKT cells, γδ Tcells, innate lymphoid cells (ILCs), macrophages, and dendritic cells(DCs) in an individual. For example, in some cases, an effective amountof an immunomodulatory composition of the present disclosure is anamount that is effective, when administered in a single dose or inmultiple doses, to modulate the number and/or activity of one or more ofNK cells, NKT cells, γδ T cells, ILCs, macrophages, and DCs in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number and/or activity of one or more ofNK cells, NKT cells, γδ T cells, ILCs, macrophages, and DCs in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate the number and/or activity of one or more of NK cells, NKTcells, γδ T cells, ILCs, macrophages, and DCs in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of NK cells, NKTcells, γδ T cells, ILCs, macrophages, and DCs in the individual in theabsence of treatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate the number and/oractivity of regulatory cells in an individual. Regulatory T cells(Tregs) are CD4⁺ or CD8⁺, and may also be FoxP3⁺ Tregs may also bedefined by other markers such as PD-1, CTLA-4 etc. Regulatory cells canalso be comprised of other innate cells such as NK, NKT, γδ T cells,ILCs and DCs, and B lymphocytes. NK and NKT can also be FoxP3⁺ and mayalso be defined by other markers such as PD-1. “Modulate the numberand/or activity” of regulatory cells, as used herein, refers toincreasing, decreasing, or balancing the number and/or activity ofregulatory cells. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate the number and/or activity of regulatory cells in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared number and/or activity of regulatory cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to increase the number of regulatory cells in an individual by at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45%, at least 50%, at least 75%, at least100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold,more than 10-fold, at least 15-fold, at least 20-fold, at least 25-fold,at least 50-fold, at least 100-fold, or more than 100-fold, compared tothe number of regulatory cells in the individual in the absence oftreatment with the immunomodulatory composition. As one example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to increase the number of CD8⁺regulatory cells (e.g., CD8⁺/CD25⁺/FoxP3⁺ cells) in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number of CD8⁺ regulatory cells (e.g., CD8⁺/CD25⁺/FoxP3⁺cells) in the individual in the absence of treatment with theimmunomodulatory composition. As another example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to increase the number of CD8⁺ regulatorycells (e.g., CD8⁺/FoxP3⁺ cells) in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber of CD8⁺ regulatory cells (e.g., CD8⁺/FoxP3⁺ cells) in theindividual in the absence of treatment with the immunomodulatorycomposition As another example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to increase the number of NKT cells (e.g., NKT⁺/FoxP3⁺ cells) in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number of NKT cells (e.g., NKT⁺/FoxP3⁺cells) in the individual in the absence of treatment with theimmunomodulatory composition. As another example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to increase the number of NKT cells (e.g.,NKT⁺/PD-1⁺ cells) in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, more than 10-fold, atleast 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, atleast 100-fold, or more than 100-fold, compared to the number of NKTcells (e.g., NKT⁺/PD-1⁺ cells) in the individual in the absence oftreatment with the immunomodulatory composition. As another example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to increase the number of NKcells (e.g., NK⁺/FoxP3⁺ cells) in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber of NK cells (e.g., NK⁺/FoxP3⁺ cells) in the individual in theabsence of treatment with the immunomodulatory composition. As anotherexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to increase thenumber of NK cells (e.g., NK⁺/PD-1⁺ cells) in an individual by at least10%, at least 15%, at least 20%, at least 25%, at least 30%, at least35%, at least 40%, at least 45%, at least 50%, at least 75%, at least100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold,more than 10-fold, at least 15-fold, at least 20-fold, at least 25-fold,at least 50-fold, at least 100-fold, or more than 100-fold, compared tothe number of NK cells (e.g., NK⁺/PD-1⁺ cells) in the individual in theabsence of treatment with the immunomodulatory composition. As anotherexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to increase thenumber of CD4⁺ regulatory cells (e.g., CD4⁺/FoxP3⁺ cells) in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number of CD4⁺ regulatory cells (e.g.,CD4⁺/FoxP3⁺ cells) in the individual in the absence of treatment withthe immunomodulatory composition. As another example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to increase the number of CD4⁺ regulatorycells (e.g., CD4⁺/CD25⁺/FoxP3⁺ cells) in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber of CD4⁺ regulatory cells (e.g., CD4⁺/CD25⁺/FoxP3⁺ cells) in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th17 cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th17 cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of Th17 cells in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific Th17 cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific Th17 cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific Th17 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of Th17 cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of Th17 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific Th17 cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific Th17 cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to normalize the level of one ormore serum markers of pathological immune responses. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, in normalizing the level of oneor more of the serum markers (these markers include, but are notrestricted to, cholesterol (CHOL), glucose (GLU), globulin (GLOB),alanine aminotransferases (ALT), aspartate aminotransferases (AST),total phosphates (TP), total bilirubin (TBIL), phosphate (PHOS),triglycerides (TRIG), uric acid (URIC), creatine kinase (CK) and urea)in an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared with the level of CH, GLU,GLOB, ALT, AST, TP, PHOS, TRIG, URIC, CK, TBIL or urea in the individualin the absence of treatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th22 cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th22 cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of Th22 cells in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific Th22 cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific Th22 cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific Th22 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of Th22 cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of Th22 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific Th22 cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the the number and/or activityof antigen-specific Th22 cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate the number and/oractivity of TH9 cells in an individual. “Modulate the number and/oractivity” of TH9 cells, as used herein, refers to increasing,decreasing, or balancing the number and/or activity of TH9 cells. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate thenumber and/or activity of TH9 cells in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared numberand/or activity of TH9 cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) and/orregulate innate and/or adaptive (including both cellular and humoral)immune responses in an individual. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of innate and/or adaptive immune cells and/or their effectorfunctions in an individual by at least 10%, at least 15%, at least 20%,at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, more than 10-fold, at least 15-fold,at least 20-fold, at least 25-fold, at least 50-fold, at least 100-fold,or more than 100-fold, compared to the number and/or activity of one ormore of innate or adaptive immune cells and/or their effector functionsin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of innate and/or adaptive immunecells and/or their effector functions in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the number and/or activity of innate and/or adaptive immunecells and/or their effector functions in the individual in the absenceof treatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to induce and/or increaseapoptosis in innate and/or adaptive immune cells in an individual toprotect from undesirable inflammation. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to induce and/or increase apoptosis in innateand/or adaptive immune cells in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber of one or more of innate or adaptive immune cells in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to induce proliferation and/ordifferentiation of hematopoietic stem cells, and restore homeostasis.For example, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to induceproliferation and/or differentiation of hematopoietic stem cells, andrestore homeostasis in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, more than 10-fold, atleast 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, atleast 100-fold, or more than 100-fold, compared to the individual in theabsence of treatment with the immunomodulatory composition.

In some cases, an immunomodulatory composition of the present disclosurecomprises CC and an antigen. Where an immunomodulatory composition ofthe present disclosure comprises CC and an antigen, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) an immune responseto the antigen by at least about 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, more than 10-fold, at least 15-fold,at least 20-fold, at least 25-fold, at least 50-fold, at least 100-fold,or more than 100-fold, compared to the immune response to the antigen inthe absence of treatment with the immunomodulatory composition. Forexample, where the antigen is an antigen associated with or derived froman autoantigen or an allergen, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) an immune response to the antigen by at leastabout 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the immune response to the antigen in the absence oftreatment with the immunomodulatory composition. For example, where theimmunomodulatory composition comprises CC, an antigen, an autoantigen oran allergen, alone or in combination with each other, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to reduce the immune response to the antigenin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the immune response tothe antigen in the individual in the absence of treatment with theimmunomodulatory composition.

The immune response can be a humoral immune response, e.g., a B cell orantibody immune response. Thus, e.g., in some cases, where the antigenis an antigen associated with or derived from an autoantigen or anallergen, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to modulate (e.g., reduce) a B cellresponse to the antigen by at least about 10%, at least 15%, at least20%, at least 25%, at least 30%, at least 35%, at least 40%, at least45%, at least 50%, at least 75%, at least 100% (or 2-fold), at least2.5-fold, at least 5-fold, at least 10-fold, more than 10-fold, at least15-fold, at least 20-fold, at least 25-fold, at least 50-fold, at least100-fold, or more than 100-fold, compared to the B cell response to theantigen in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the B cell response to the antigen in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the B cell response to the antigen in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, where the antigen is an antigenassociated with or derived from an autoantigen or an allergen, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the amount ofantibody specific to the antigen by at least about 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, more than 10-fold, atleast 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, atleast 100-fold, or more than 100-fold, compared to the amount ofantibody specific to the antigen in the absence of treatment with theimmunomodulatory composition. For example, in some cases, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to reduce the amount of antibody specific to the antigenin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the amount of antibodyspecific to the antigen in the individual in the absence of treatmentwith the immunomodulatory composition.

The immune response can be a cellular immune response, e.g., a T cellresponse. Thus, e.g., in some cases, where the antigen is an antigenassociated with or derived from an autoantigen or an allergen, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate a (e.g., reduce) T cell responseto the antigen by at least about 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, more than 10-fold, at least 15-fold,at least 20-fold, at least 25-fold, at least 50-fold, at least 100-fold,or more than 100-fold, compared to the T cell response to the antigen inthe absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce the Tcell response to the antigen in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the T cell response to the antigen in the individual in theabsence of treatment with the immunomodulatory composition. In somecases, the immune response is a humoral immune response and a cellularimmune response.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10³ CC per ml to about 10¹² CC per ml. Forexample, an immunomodulatory composition of the present disclosure cancomprise CC in an amount of from about 10³ CC per ml to about 10⁴ CC perml, from about 10⁴ CC per ml to about 10⁵ CC per ml, from about 10⁵ CCper ml to about 10⁶ CC per ml, from about 10⁶ CC per ml to about 10⁷ CCper ml, from about 10⁸ CC per ml to about 10⁹ CC per ml, from about 10⁹CC per ml to about 10¹⁰ CC per ml, from about 10¹⁶ CC per ml to about10¹¹ CC per ml, or from about 10¹¹ CC per ml to about 10¹² CC per ml.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10³ CC per mg to about 10¹² CC per mg. Forexample, an immunomodulatory composition of the present disclosure cancomprise CC in an amount of from about 10³ CC per mg to about 10⁴ CC permg, from about 10⁴ CC per mg to about 10⁵ CC per mg, from about 10⁵ CCper mg to about 10⁶ CC per mg, from about 10⁶ CC per mg to about 10⁷ CCper mg, from about 10⁸ CC per mg to about 10⁹ CC per mg, from about 10⁹CC per mg to about 10¹⁰ CC per mg, from about 10¹⁶ CC per mg to about10¹¹ CC per mg, or from about 10¹¹ CC per mg to about 10¹² CC per mg.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10³ CC per gram to about 10¹⁵ CC per gram.For example, an immunomodulatory composition of the present disclosurecan comprise CC in an amount of from about 10³ CC per gram to about 10⁴CC per gram, from about 10⁴ CC per gram to about 10⁵ CC per gram, fromabout 10⁵ CC per gram to about 10⁶ CC per gram, from about 10⁶ CC pergram to about 10⁷ CC per gram, from about 10⁸ CC per gram to about 10⁹CC per gram, from about 10⁹ CC per gram to about 10¹⁰ CC per gram, fromabout 10¹⁰ CC per gram to about 10¹¹ CC per gram, from about 10¹¹ CC pergram to about 10¹² CC per gram, from about 10¹² CC per gram to about10¹³ CC per gram, from about 10¹³ CC per gram to about 10¹⁴ CC per gram,or from about 10¹⁴ CC per gram to about 10¹⁵ CC per gram.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10² to about 10²⁰ colony forming units(cfu) per unit dosage form; for example, an immunomodulatory compositionof the present disclosure can comprise CC in an amount of from about 10²to about 10³ from about 10³ to about 10⁵, from about 10⁵ to about 10⁷,from about 10⁷ to about 10⁹, from about 10⁹ to about 10¹¹, from about10¹¹ to about 10¹³, from about 10¹³ to about 10¹⁵, from about 10¹⁵ toabout 10¹⁸, or from about 10¹⁸ to about 10²⁰, cfu per unit dosage form.A unit dosage form can be an amount that is administered in a singledose; for example, a unit dosage form can be 0.5 ml, 1.0 ml, or othervolume suitable for administration in a single dose.

CC can be prepared by exposing Caulobacter crescentus to a temperatureof from about 0° C. to about 37° C. (e.g., from about 0° C. to 15° C.;from about 10° C. to 20° C.; from about 20° C. to 25° C.; from about 23°C. to 25° C.; from about 23° C. to 37° C.; or from about 30° C. to 35°C.) for a time period of from about 1 hour to extended periods of time(e.g., at least 1 hour; at least 2 hours; at least 4 hours; overnight;at least 24 hours; at least 48 hours; at least 100 hours, or more than100 hours). CC can also be stored in saline, phosphate-buffered saline(PBS), or any other buffer, at temperatures from about 36° C. to −170°C. (e.g., from about 0° C. to 36° C.; from about 0° C. to 4° C.; fromabout 10° C. to 15° C.; from about 0° C. to −20° C.; from about −10° C.and below), or in conditions known to those skilled in the art. CC canbe 0.0000001 to 100% viable. For example, CC can be 0.0000001 to0.000001% viable, 0.000001 to 0.00001% viable, 0.00001 to 0.0001%viable, 0.0001% to 0.001% viable, 0.001% to 0.01% viable, 0.01% to 0.1%viable, 0.1% to 1% viable, 1% to 10% viable, from 10% to 100% viable,from 25% to 100 viable, from 50% to 100% viable, from 75% to 100%viable, or from 90% to 100% viable.

An immunomodulatory composition of the present disclosure can compriseCC grown in culture at various optical density units (ODs) from about0.1 OD to 30.0 ODs. For example, the OD of the CC culture grown can be0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 etc.

Caulobacter crescentus

An immunomodulatory composition of the present disclosure comprisesCaulobacter, where the Caulobacter is non-pathogenic. The non-pathogenicCaulobacter genus includes 19 different species, including two speciesof Asticcacaulis (C. vibroides, C. henricii, C. intermedius, C.robiginosus, C. rutilis, C. subvibriodes, C. fusiformis, C. rossii, A.excentricus, A. biprosthecum etc.). See, e.g., JS Poindexter, TheCaulobacters: Ubiquitous Unusual Bacteria, Microbiol Rev 45, 123-179,1981). Several of the Caulobacter sp. are available from the AmericanType Culture Collection (ATCC), such as CB35, CB26, CB28, KAS, CB66, FC4etc. All of these species of Caulobacter in live, non-denatured, mutatedor attenuated forms can be used as immunomodulatory agents describedherein. In addition, Caulobacter bacteria can be in non-motileprosthecate, motile swarmer, stubby flagellin and flagellin positive,flagellin negative, dividing and/or non-dividing forms. Caulobacter sp.can be grown at temperatures ranging from 18°-42° C., and pH rangingfrom 5-9, but optimally at a temperature in a range of 23−25° C. and pH7 in PYE medium. Mutated or genetically modified forms of Caulobactersp. can be produced by modifying the nutrients, chemicals, pH,temperature, ultraviolet or infrared light, radiation etc. of theculture conditions, or genetically modifying various enzymes, metabolicpathways, surface molecules, nucleic acids, plasmids, cellular and cellwall components, smooth and rough LPS in live bacteria (JS Poindexter,The Caulobacters: Ubiquitous Unusual Bacteria, Microbiol Rev 45,123-179, 1981).

Caulobacter crescentus can act as a carrier and/or delivery vehicle todeliver antigens. As a non genetic modification (GM), such aselectrostatic and hydrophobic interactions, binding of antigens to theCaulobacter crescentus surface may enable the Caulobacter crescentus toact as an antigen carrier and/or delivery vehicle. Further, due tobioadhesion/mucoadhesion, Caulobacter crescentus may facilitate antigenuptake by M cell transport, delivery to and subsequent modulation ofDCs/APCs, modulation of NK, NKT, B and T cell responses at mucosalsurfaces.

Although the discussion below focuses on Caulobacter crescentus, any ofa variety of non-pathogenic Caulobacter species can be included in animmunomodulatory composition of the present disclosure.

In some cases, an immunomodulatory composition of the present disclosurecomprises Caulobacter crescentus (CC). In some cases, the Caulobactercrescentus is wild-type. In some cases, the Caulobacter crescentus is alipopolysaccharide-negative strain. In some cases, the Caulobactercrescentus is an S-layer-negative strain. In some cases, the CC ismutated attenuated, or contains suicidal mutations. In some cases,Caulobacter crescentus is with or without a drug resistant plasmid suchas chloramphenicol, penicillin resistant plasmids. In some cases,Caulobacter crescentus can be grown in other medium than PYE medium.

In some cases, the Caulobacter crescentus is genetically modified toproduce one or more heterologous polypeptides. The polypeptides can beof a wide range of sizes. Suitable heterologous polypeptides include,but are not limited to, PD1, PDL, CTLA-4, GITR, VISTA; a co-inhibitoryprotein found on antigen-presenting cells (APCs) or T cells; a cytokine(e.g., IL-10; or any of the above-listed cytokines); a chemokine; anantigen (e.g., an autoantigen or allergen as described herein above); anantibody against an antigen (e.g., an autoantigen; as described hereinabove), a signalling molecule, a receptor, a cytokine, a pro-apoptoticprotein; a fusion protein (e.g., an antigen and a cytokine, an antigenand a carrier protein) etc.

In some cases, Caulobacter crescentus is modified by labeling orcoupling the bacterium with fluorescent, radioactive isotope, light tagsetc.

In some cases, Caulobacter crescentus is genetically modified. In somecases, Caulobacter crescentus is genetically modified so that microbe isattenuated. In some cases, the nucleic acid of the Caulobactercrescentus is modified so that the microbe is attenuated forproliferation.

In some cases, an immunomodulatory composition of the present disclosurecomprises whole CC. In some cases, an immunomodulatory composition ofthe present disclosure comprises CC that are live or non-denatured. Insome cases, an immunomodulatory composition of the present disclosurecomprises individual or multiple components, byproducts and/ormetabolites of CC, which can be isolated, synthesized, or geneticallymanufactured in other synthetic or natural bacterial cell as syntheticbiotics. The components of CC can comprise, but are not limited to,effector molecules e.g., polysaccharides, glycosylceramides,peptidoglycans, nucleic acids, structural proteins, short chain fattyacids, fatty acid metabolites, hydroxyl fatty acids etc. Fractions,components, by-products and/or metabolites of Caulobacter crescentus canbe obtained by filtering culture supernatants, treatment with variousorganic solvents, enzymes such as glycosidases, lipase, DNAse, RNAse,protease, lysozyme etc. In some cases, an immunomodulatory compositionof the present disclosure comprises individual or multiple components ofCC, which can be fused with antigens using physicochemical or geneticmethods and used as synthetic bacteria.

In some cases, Caulobacter crescentus is bioengineered in its outermembrane vesicle to package and deliver chemotherapeutics and/orimmunotherapeutics and synthetic, genetic material from other bacteria.In some cases, Caulobacter crescentus is bioengineered and constructedinto a genetic circuit as a synthetic therapeutic bacteria, “syntheticbiotics”.

In some cases, an immunomodulatory composition of the present disclosurecomprises S-layer of CC. In some cases, an immunomodulatory compositionof the present disclosure comprises S-layer of CC that is geneticallymodified to display one or more heterologous polypeptides,chemotherapeutics and/or immunotherapeutics. In some cases, animmunomodulatory composition of the present disclosure comprisescomponents of S-layer.

Antigens

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, ormore than 10) antigens. Suitable antigens include, but are not limitedto, an antigen derived from an autoantigen; and an allergen.

In some embodiments, Caulobacter crescentus is genetically modified toproduce an antigen; and the genetically modified Caulobacter crescentusis live, to produce an immunomodulatory composition of the presentdisclosure. Methods of genetically modifying bacteria are known in theart.

In other embodiments, CC is admixed with an antigen in animmunomodulatory composition of the present disclosure. Caulobactercrescentus can act as a carrier and/or delivery vehicle to deliverantigens. As a non genetic modification (GM), such as electrostatic andhydrophobic interactions, binding of antigens to the Caulobactercrescentus surface may enable the Caulobacter crescentus to act as anantigen carrier and/or delivery vehicle. Further, due tobioadhesion/mucoadhesion, Caulobacter crescentus may facilitate antigenuptake by M cell transport, delivery to and subsequent modulation ofDCs/APCs, modulation of NK, NKT, B and T cell responses at mucosalsurfaces.

An antigen, for use in certain embodiments of the herein describedimmunomodulatory compositions and methods employing CC, may be anytarget epitope, molecule, molecular complex, cell or tissue againstwhich modulation of immunogenicity in a subject is desired.

An immunomodulatory composition of the present disclosure can includeone or more antigens or antigenic compositions capable of modulating animmune response against a human or animal autoantigen or allergen. Theantigen may be associated with autoimmune disease, allergy, asthma,prion disease or any other conditions where modulation of anantigen-specific response would be desirable or beneficial.

A suitable antigen can be any type of antigen known in the art. Antigenscan be produced in any of a variety of sources such as plants, animals,prokaryotes, in vitro cell culture, etc. Antigens can be in variety offorms as described below.

Suitable antigens include, e.g., peptides, modified peptides, peptidemimotopes, conformationally-constrained synthetic peptides,multi-epitope peptides from one or more antigens, branched peptides,lipopeptides, monolipopeptides, dilipopeptides, peptides conjugated orfused to proteins, peptides conjugated or fused to T cell or B cellepitopes. See, e.g., U.S. Pat. No. 8,198,400. Suitable antigens include,e.g., full-length antigens, truncated antigens, mutated antigens, andinactivated or combined forms. Suitable antigens include, e.g.,proteins, purified or recombinant proteins, recombinant fusion proteins,proteins and peptides conjugated to toll-like receptor (TLR)agonists/antagonists, proteins and peptides conjugated to bacterialtoxins, proteins and peptides conjugated to antibodies, proteins andpeptides conjugated to cytokines and chemokines, glycoproteins,glycolipoproteins and derivatives thereof. Suitable antigens include,e.g., polysaccharides, polysaccharide conjugates, oligosaccharides,lipids, glycolipids, carbohydrates and derivatives thereof. An antigencan be modified to modulate antigen presentation and/or co-inhibition,or increase co-inhibitory signals.

An antigen or antigenic composition can be obtained from autoantigens,allergens, etc.

An antigen can be a whole cell extract, a cell lysates, a whole cell, awhole live cell, a whole inactivated cell, a whole irradiated cell, etc.Antigens may be crude, purified, or recombinant form. In some cases, anantigen is at least 50% pure, at least 60% pure, at least 70% pure, atleast 80% pure, at least 90% pure, at least 95% pure, at least 98% pure,or at least 99% pure, or more than 99% pure.

An antigen can be chemically, enzymatically, or genetically coupled toCC. In some cases, an antigen is present in an immunomodulatorycomposition of the present disclosure in admixture with CC.

An immunomodulatory composition of the present disclosure can comprise asingle type of antigen. An immunomodulatory composition of the presentdisclosure can include 2 or more different antigens. An immunomodulatorycomposition of the present disclosure can include 2, 3, 4, 5, 6, or morethan 6, different antigens. Where an immunomodulatory composition of thepresent disclosure includes more than one antigen, the more than oneantigen can be from the same cell or allergen. Where an immunomodulatorycomposition of the present disclosure includes more than one antigen,the more than one antigen can be from two or more cells, or allergens.

An antigen can be in the form of a protein, a lipopolysaccharide, alipoprotein, a proteoglycan, glycoproteins, glycosaminoglycans, anoligosaccharide, etc.

An antigen can be in the form of a nucleic acid comprising a nucleotidesequence encoding an antigen, e.g., a polypeptide antigen. For example,an antigen can be provided in the form of DNA (e.g., plasmid DNA, nakedDNA etc.), RNA, and/or a wild-type, attenuated and/or recombinantvector-based nucleic acid. The nucleic acid coding for the antigen canbe either “naked” or contained in a delivery system, such as liposomes.

A recombinant vector-encoded antigen can be at least one recombinantexpression construct which comprises a promoter operably linked to anucleotide sequence encoding an antigen in recombinant viral vectors(such as adenovirus (e.g. Ad2, Ad4, Ad5, Ad35, Ad35K5 etc.),adeno-associated virus, lentivirus, herpes virus, poxvirus, vesicularstomatitis virus, alpha virus, measles virus, papaya mosaic virus,cytomegalovoirus, modified vaccinia Ankara virus MVA, polio virus, Marbavirus etc.), bacterial vector vaccines (such as Salmonella, Shigella, E.coli, Lactococcus lactic, Listeria sp., Lactobacillus sp.), fungalvectors (such as heat killed recombinant Saccharomyces yeast), plantviruses, virus-like particles (VLPs), virosomes, synthetic vaccineparticles, synthetic biomimetic supramolecular biovectors,depathogenized viral/bacterial strains (such as NIBRG14 from H5N1). Thevector could be in the form of live wild-type, non-replicative, mutated,modified, defective or attenuated. The vectors could be from human,animal, plant or prokaryote origin and in any effective amount.

In treating or preventing autoimmune diseases or allergy, antigen can begiven at the same or different times, at the same or different site thanthe immunomodulatory composition of the present disclosure.

Autoantigens

In some cases, an immunomodulatory composition of the present disclosurecomprises, in addition to CC, an autoantigen. In some cases, animmunomodulatory composition of the present disclosure comprises, inaddition to CC, one or more autoantigens, e.g., 1, 2, 3, 4, 5, or moreantigens, from one or more self tissues.

For example, where the autoimmune disease is type 1 diabetes, an antigencan be pancreatic islet beta cell associated antigen, proinsulin,glutamic acid decarboxylase, chromogranin A, islet amyloid polypeptide,HSP60; for systemic lupus erythematosus, an antigen can be snRNP; forGrave's disease, an antigen can be thyroglobulin, thyrotropin receptoror a thyroid epithelial cell; for thrombocytopenic purpura, an antigencan be a platelet, GPIIB/IIIa; for multiple sclerosis, an antigen can bemyelin basic protein, MOG, PLP; for celiac disease, an antigen can betransglutaminidase.

A suitable autoantigen can be an autoantigen involved in the initiationand/or propagation of an autoimmune disease, the pathology of which canbe due to the presence of antibodies specific for a molecule expressedby the relevant target organ, tissue, or cells, e.g., systemic lupuserythematosus (SLE) or myasthenia gravis (MG). In such diseases, it canbe desirable to direct an ongoing antibody-mediated (i.e., a Th2-type)immune response to the relevant autoantigen towards a cellular (i.e., aTh1-type) immune response. Alternatively, it can be desirable to preventonset of or decrease the level of a Th2 response to the autoantigen in asubject not having, but who is suspected of being susceptible to, therelevant autoimmune disease by prophylactically inducing a Th1 responseto the appropriate autoantigen. Autoantigens that can be included in asubject immunomodulatory composition include, without limitation: (a)with respect to SLE, the Smith protein, RNP ribonucleoprotein, and theSS-A and SS-B proteins; and (b) with respect to MG, the acetylcholinereceptor. Examples of other antigens involved in one or more types ofautoimmune response include, e.g., endogenous hormones such asluteinizing hormone, follicular stimulating hormone, testosterone,growth hormone, prolactin, and other hormones.

Other examples of suitable autoantigens include antigens associated withneurological diseases such as schizophrenia, Alzheimer's disease,depression, hypopituitarism, and cardiovascular diseases such asatherosclerosis (e.g., an antigen for atherosclerosis can be cholesterylester transfer protein, oxidized LDL, apoB210, apoB100) etc.

Those of skill in the art will recognize that other suitableautoantigens include those that are associated with juvenile rheumatoidarthritis and Marie-Strumpell ankylosing spondylitis, that can lead toanterior uveitis and subsequent glaucoma. Other suitable autoantigensinclude those that are associated with Huntington's disease, andParkinson's disease.

Examples of autoantigens include those that are associated with cell ororgan-specific autoimmunity. Such autoantigens include the acetylcholinereceptor, associated with Myasthenia gravis; actin, associated withchronic active hepatitis and primary biliary cirrhosis; adeninenucleotide translocator (ANT), associated with dilated cardiomyopathyand myocarditis; β-adrenoreceptor, associated with dilatedcardiomyopathy; aromatic L-amino acid decarboxylase, associated withautoimmune polyendocrine syndrome type I (APS-I); asialoglycoproteinreceptor, associated with autoimmune hepatisis;bactericidal/permeability-increasing protein (Bpi), associated withcystic fibrosis vasculitides; calcium-sensing receptor, associated withacquired hypoparathryoidism; cholesterol side-chain cleavage enzyme(CYPIIa), associated with APS-I; collagen type IV α₃-chain; associatedwith Goodpasture syndrome; cytochrome P450 2D6 (CYP2D6), associated withautoimmune hepatisis; desmin, associated with Crohn disease and coronaryartery disease; desmoglein 1, associated with pemphigus foliaceus;desmoglein 3, associated with pemphigus vulgaris; F-actin, associatedwith autoimmune hepatitis; GM gangliosides, associated withGuillain-Barré syndrome; glutamate decarboxylase (GAD65), associatedwith type 1 diabetes and stiff man syndrome; glutamate receptor (GLUR),associated with Rasmussen encephalitis; H/K ATPase, associated withautoimmune gastritis; 17-α-hydroxylase (CYP17), associated with APS-I;21-hydroxylast (CYP21), associated with Addison disease; IA-2 (ICA512),associated with type 1 diabetes; insulin, associated with type 1diabetes and insulin hypoglycemic syndrome (Hirata disease); insulinreceptor, associated with type B insulin resistance, acanthosis andsystemic lupus erythematosus (SLE); intrinsic factor type 1, associatedwith pernicious anemia, leukocyte function-associated antigen (LFA-1),associated with treatment-resistant Lyme arthritis; myelin-associatedglycoprotein (MAG), associated with polyneuropathy; myelin basicprotein, associated with multiple sclerosis and demyelinating diseases;myelin oligodendrocyte glycoprotein (MOG), associated with multiplesclerosis; myosis, associated with rheumatic fever; p-80-coilin,associated with atopic dermatitis; pyruvate dehydrogenase complex-E2(PDC-E2), associated with primary biliary cirrhosis; sodium iodidsymporter (NIS), associated with Graves disease and autoimmunehypothyroidism; SOX-10, associated with vitiligo; thyroid and eye muscleshared protein, associated with thyroid associated ophthalmopathy;thyroglobulin, associated with autoimmune thyroiditis; thyroidperoxidase, associated with autoimmune Hashimoto thyroiditis;thyrotropin receptor, associated with Graves disease; tissuetransglutaminase, associated with coeliac disease; transcriptioncoactivator p75, associated with atopic dermatitis; tryptophanhydroxylase, associated with APS-I; tyrosinase, associated with vitiligoand metastatic melanoma; and tyrosine hydroxylase, associated withAPS-I.

Examples of autoantigens include those that are associated with systemicautoimmunity. Such autoantigens include ACTH, associated with ACTHdeficiency; aminoacyl-tRNA histidyl synthetase, associated with myotitisand dermatomyositis; aminoacyl-tRNA synthetase (several), associatedwith polymyositis and dermatomyositis; cardiolipin, associated with SLE;carbonic anhydrase II, associated with SLE, Sjögren syndrome andsystemic sclerosis; collagen (multiple types), associated withrheumatoid arthritis (RA), SLE and progressive systemic sclerosis;centromere-associated proteins, associated with systemic sclerosis;DNA-dependent nucleosome-stimulated ATPase, associated withdermatomyositis; fibrillarin, associated with scleroderma; fibronectin,associated with SLE, RA and morphea; glucose-6-phosphate isomerase,associated with RA; β2-glycoprotein I (β2-GPI), associated with primaryantiphospholipid syndrome; golgin (95, 97, 160, 180), associated withSjögren syndrome, SLE and RA; heat shock protein, associated withvarious immune-related disorders; hemidesmosomal protein 180, associatedwith bullous pemphigoid, herpes gestationis and cicatricial pemphigoid;histone H2A-H2B-DNA, associated with SLE; IgE receptor, associated withchronic idiopathic urticaria; keratin, associated with RA;Ku-DNA-protein kinase, associated with SLE; Ku-nucleoprotein, associatedwith connective tissue syndromes; La phosphoprotein (La 55-B),associated with Sjögren syndrome; myeloperoxidase, associated withnecrotizing and crescentic glomerulonephritis and systemic vasculitis;proteinase 3 (PR3), associated with Wegener granulomatosis andChurg-Strauss syndrome; RNA polymerase I-III (RNP), associated withsystemic sclerosis and SLE; signal recognition protein (SRP54),associated with polymyositis; topoisomerase-I (Scl-70), associated withscleroderma and Raynaud syndrome; tubulin, associated with chronic liverdisease and visceral leishmaniasis; and vimentin, associated withsystemic autoimmune disease.

Other examples of autoantigens include those that are associated withplasma protein and cytokine autoimmunity Such autoantigens include C1inhibitor, associated with autoimmune C1 deficiency; C1q, associatedwith SLE and membrane proliferative glomerulonephritis (MPGN); cytokines(IL-1α, IL-1β, IL-6, IL-10, LIF), associated with RA and systemicsclerosis; factor II, factor V, factor VII, factor VIII, factor IX,factor X, factor XI, factor XII, thrombin, vWF, associated withprolonged coagulation time; glycoprotein IIb/IIIg and 1B/IX, associatedwith autoimmune thrombocytopenia purpura; IgA, associated withimmunodeficiency; and oxidized LDL (OxLDL), associated withartherosclerosis.

Yet more examples of autoantigens include those that are associated withcancer and paraneoplastic autoimmunity. Such autoantigens includeamphiphysin, associated with neuronopathy and small lung cell cancer;cyclin B1, associated with hepatocellular carcinoma; DNA topoisomeraseII, associated with liver cancer; desmoplakin, associated withparaneoplastic pemphigus; gephyrin, associated with paraneoplastic stiffman syndrome; Hu proteins, associated with paraneoplasticencephalomyelitis; neuronal nicotinic acetylcholine receptor, associatedwith subacute autonomic neuropathy and cancer; p53, associated withcancer and SLE; p62 (IGF-II mRNA-binding protein), associated withhepatocellular carcinoma (China); recoverin, associated withcancer-associated retinopathy; Ri protein, associated withparaneoplastic opsoclonus myoclonus ataxia; β IV spectrin, associatedwith lower motor neuron syndrome; synaptotagmin, associated withLambert-Eaton myasthenic syndrome; voltage-gated calcium channels,associated with Lamber-Eaton myasthenic syndrome; and Yo protein,associated with paraneoplastic cerebellar degeneration.

Allergens

In some cases, an immunomodulatory composition of the present disclosurecomprises, in addition to CC, an allergen. Suitable allergens can beobtained and/or produced using known methods. Classes of suitableallergens include, but are not limited to, pollens, animal dander otherthan cat dander, grasses, molds, dusts, antibiotics, stinging insectvenoms, and a variety of environmental (including chemicals and metals),drug and food allergens. Common tree allergens include pollens fromcottonwood, popular, ash, birch, maple, oak, elm, hickory, and pecantrees; common plant allergens include those from mugwort, ragweed,English plantain, sorrel-dock and pigweed; plant contact allergensinclude those from poison oak, poison ivy and nettles; common grassallergens include rye grass, Timothy, Johnson, Bermuda, fescue andbluegrass allergens; common allergens can also be obtained from molds orfungi such as Alternaria, Fusarium, Hormodendrum, Aspergillus,Micropolyspora, Mucor and thermophilic actinomycetes; epidermalallergens can be obtained from house or organic dusts (typically fungalin origin), from arthropods such as house mites (Dermatophagoidespteronyssinus), or from animal sources such as feathers, and dog dander;common food allergens include milk and cheese (diary), egg, wheat, nut(e.g., peanut), seafood (e.g., shellfish), pea, bean and glutenallergens; common environmental allergens include metals (nickel andgold), chemicals (formaldehyde, trinitrophenol and turpentine), Latex,rubber, fiber (cotton or wool), burlap, hair dye, cosmetic, detergentand perfume allergens; common drug allergens include local anestheticand salicylate allergens; antibiotic allergens include penicillin,tetracycline and sulfonamide allergens; and common insect allergensinclude bee, wasp and ant venom, and cockroach calyx allergens.Particularly well characterized allergens include, but are not limitedto, the major and cryptic epitopes of the Der p I allergen (Hoyne et al.(1994) Immunology 83190-195), bee venom phospholipase A2 (PLA) (Akdis etal. (1996) J. Clin. Invest. 98:1676-1683), birch pollen allergen Bet v 1(Bauer et al. (1997) Clin. Exp. Immunol. 107:536-541), and themulti-epitopic recombinant grass allergen rKBG8.3 (Cao et al. (1997)Immunology 90:46-51). These and other suitable allergens arecommercially available and/or can be readily prepared as extractsfollowing known techniques.

Suitable allergens include tree pollen allergens, weed pollen allergens,herb pollen allergens, grass pollen allergens, mite allergens, insectallergens, venom allergens, animal hair allergens, dander allergens andfood allergens.

In some cases, the allergen is in the form of an extract, a purifiedallergen, a modified allergen or a recombinant allergen or a mutant of arecombinant allergen or any combination thereof. In some cases, theallergen is selected from the group consisting of grass pollen allergen,dust mite allergen, ragweed allergen, cat allergen and birch allergen.

An allergen can be present in an immunomodulatory composition of thepresent disclosure in an amount of from about 2.5 μg to about 75 μg perunit dosage form. For example, an allergen can be present in animmunomodulatory composition of the present disclosure in an amount offrom about 2.5 μg to about 5 μg, from about 5 μg to about 10 ng, fromabout 10 μg to about 15 ng, from about 15 μg to about 20 μg, from about20 μg to about 25 μg, from about 25 μg to about 50 μg, or from about 50μg to about 75 μg, or more than 75 μg, per unit dosage form.

In some cases, a dose of an immunomodulatory composition of the presentdisclosure that comprises an allergen has a potency of about 65 to about17,600 Biological Allergen Units (BAU). In some cases, a dose of animmunomodulatory composition of the present disclosure that comprises anallergen comprises from about 650 BAU to about 6,000 BAU.

Antibodies

In some cases, an immunomodulatory composition of the present disclosurecomprises, in addition to CC, an antibody against a cancer antigen, anautoantigen, an allergen or a pathogenic antigen (e.g., a therapeuticantibody, monoclonal antibodies, bispecific antibodies, chemoimmunoconjugated antibodies, radioimmunoconjugated antibodies,antibody-cytokine fusion proteins, antibody-antigen fusion proteins,antibody-immunotoxin fusion protein etc.).

Antibodies that can be included in an immunomodulatory composition ofthe present disclosure include, without limitation, antibodies directedagainst co-stimulatory or co-inhibitory molecules (CD28, CD40, CTLA-4,PD-1, PDL-1, GITR, VISTA, LAG-3, ICOS, CD137, OX40, CD137, CD227,CTLA-4, KIRs, TCR, TIM3 etc.); and other therapeutic antibodies.

Non-limiting examples of suitable antibodies include, but are notlimited to, adalimumab, bevacizumab, infliximab, abciximab, alemtuzumab,bapineuzumab, basiliximab, belimumab, briakinumab, brodalumab,canakinumab, certolizumab pegol, cetuximab, conatumumab, denosumab,eculizumab, etrolizumab, gemtuzumab ozogamicin, golimumab, ibritumomabtiuxetan, labetuzumab, mapatumumab, matuzumab, mepolizumab, motavizumab,muromonab-CD3, natalizumab, nimotuzumab, ofatumumab, omalizumab,oregovomab, palivizumab, panitumumab, pemtumornab, pertuzumab,ranibizumab, rituximab, rovelizumab, tocilizumab, tositumomab,trastuzumab, ustekinumab, vedolizomab, zalutumumab, and zanolimumab.

Non-limiting examples of therapeutic and prophylactic antibodies thatcan be used in combination with an immunomodulatory composition of thepresent disclosure include MDX-010 (Medarex, N.J.) which is a humanizedanti-CTLA-4 antibody for the treatment of prostate cancer; SYNAGIS™(MedImmune, Md.) which is a humanized anti-respiratory syncytial virus(RSV) monoclonal antibody for the treatment of RSV infection; andHERCEPTIN™ (Trastuzumab) (Genentech, Calif.) which is a humanizedanti-HER2 monoclonal antibody for the treatment of metastatic breastcancer. Other examples are humanized anti-CD18 F(ab′)₂ (Genentech);CDP860 which is a humanized anti-CD18 F(ab′)₂ (Celltech, UK); PRO542which is an anti-HIV gp120 antibody fused with CD4 (Progenics/GenzymeTransgenics); Ostavir which is a human anti-Hepatitis B virus antibody(Protein Design Lab/Novartis); PROTOVIR™ which is a humanized anti-CMVIgGI antibody (Protein Design Lab/Novartis); MAK-195 (SEGARD) which is amurine anti-TNF-α F(ab′)₂ (Knoll Pharma/BASF); IC14 which is ananti-CD14 antibody (ICOS Pharm); a humanized anti-VEGF IgG1 antibody(Genentech); OVAREX™ which is a murine anti-CA 125 antibody (Altarex);PANOREX™ which is a murine anti-17-IA cell surface antigen IgG2aantibody (Glaxo Wellcome/Centocor); BEC2 which is a murine anti-idiotype(GD3 epitope) IgG antibody (ImClone System); IMC-C225 which is achimeric anti-EGFR IgG antibody (ImClone System); VITAXIN™ which is ahumanized anti-αVβ3 integrin antibody (Applied MolecularEvolution/MedImmune); Campath 1H/LDP-03 which is a humanized anti-CD52IgG1 antibody (Leukosite); Smart M195 which is a humanized anti-CD33 IgGantibody (Protein Design Lab/Kanebo); RITUXAN™ which is a chimericanti-CD20 IgG1 antibody (IDEC Pharm/Genentech, Roche/Zettyaku);LYMPHOCIDE™ which is a humanized anti-CD22 IgG antibody (Immunomedics);Smart ID10 which is a humanized anti-HLA antibody (Protein Design Lab);ONCOLYM™ (Lym-1) is a radiolabelled murine anti-HLA DIAGNOSTIC REAGENTantibody (Techniclone); ABX-IL8 is a human anti-IL8 antibody (Abgenix);anti-CD11a is a humanized IgG1 antibody (Genentech/Xoma); ICM3 is ahumanized anti-ICAM3 antibody (ICOS Pharm); IDEC-114 is a primatizedanti-CD80 antibody (IDEC Pharm/Mitsubishi); ZEVALIN™ is a radiolabelledmurine anti-CD20 antibody (IDEC/Schering AG); IDEC-131 is a humanizedanti-CD40L antibody (IDEC/Eisai); IDEC-151 is a primatized anti-CD4antibody (IDEC); IDEC-152 is a primatized anti-CD23 antibody(IDEC/Seikagaku); SMART anti-CD3 is a humanized anti-CD3 IgG (ProteinDesign Lab); 5G1.1 is a humanized anti-complement factor 5 (C5) antibody(Alexion Pharm); D2E7 is a humanized anti-TNF-α antibody (CAT/BASF);CDP870 is a humanized anti-TNF-α Fab fragment (Celltech); IDEC-151 is aprimatized anti-CD4 IgG1 antibody (IDEC Pharm/SmithKline Beecham);MDX-CD4 is a human anti-CD4 IgG antibody (Medarex/Eisai/Genmab); CDP571is a humanized anti-TNF-α IgG4 antibody (Celltech); LDP-02 is ahumanized anti-α4β7 antibody (LeukoSite/Genentech); OrthoClone OKT4A isa humanized anti-CD4 IgG antibody (Ortho Biotech); ANTOVA™ is ahumanized anti-CD40L IgG antibody (Biogen); ANTEGREN™ is a humanizedanti-VLA-4 IgG antibody (Elan); MDX-33 is a human anti-CD64 (FcγR)antibody (Medarex/Centeon); SCH55700 is a humanized anti-IL-5 IgG4antibody (Celltech/Schering); SB-240563 and SB-240683 are humanizedanti-IL-5 and IL-4 antibodies, respectively, (SmithKline Beecham);rhuMab-E25 is a humanized anti-IgE IgG1 antibody(Genentech/Norvartis/Tanox Biosystems); ABX-CBL is a murine anti CD-147IgM antibody (Abgenix); BTI-322 is a rat anti-CD2 IgG antibody(MedImmune/Bio Transplant); Orthoclone/OKT3 is a murine anti-CD3 IgG2aantibody (ortho Biotech); SIMULECT™ is a chimeric anti-CD25 IgG1antibody (Novartis Pharm); LDP-01 is a humanized anti-β₂-integrin IgGantibody (LeukoSite); Anti-LFA-1 is a murine anti CD18 F(ab′)₂(Pasteur-Merieux/Immunotech); CAT-152 is a human anti-TGF-β₂ antibody(Cambridge Ab Tech); and Corsevin M is a chimeric anti-Factor VIIantibody (Centocor). The above-listed immunoreactive reagents, as wellas any other immunoreactive reagents, may be administered according toany regimen known to those of skill in the art, including the regimensrecommended by the suppliers of the immunoreactive reagents.

Other examples of therapeutic and prophylactic antibodies that can beused in combination with an immunomodulatory composition of the presentdisclosure include Humira and Remicade; ACTEMRA™ (Genentech) which is arecombinant monoclonal IgG1 anti-human interleukin 6-receptor antibodyfor the treatment of anti-TNF resistant rheumatoid arthritis (RA) andjuvenile idiopathic arthritis (JIA); ARZERRA™ (GlaxoSmithKline/Novartis)which is a chimeric human monoclonal antibody directed against membraneproximal epitope on the CD20 molecule for the treatment of RA; BENLYSTA™(GlaxoSmithKline) which is a human monoclonal IgG1 gamma that binds toand inhibits the soluble form of the B-lymphocyte stimulator (BLyS)protein for the treatment of SLE; ORENCIA™ (Bristol-Myers Squibb) whichis a CTLA-4 IgG1 binding to CD80/86 on antigen-presenting cellsinhibiting the co-stimulation of CD28 on the T cells for the treatmentof RA, JIA and SLE; SIMPONI (Janssen) which is a IgG1 monoclonalantibody acting on both soluble and membrane-bound TNF-α for thetreatment of RA, psoriatic arthritis (PsA) and ankylosing spondylitis(AS); CIMZIA™ (UCB Group) which is a pegylated humanized antibody Fab′fragment of the TNF-α monoclonal antibody for the treatment of RA;Sifalimumab (MedImmune) which is an anti-IFN-α monoclonal antibodydesigned for the treatment of SLE, dematomyositis and polymyositis;various intravenous immunoglobulin products which are pools ofimmunoglobulins from healthy individuals for the treatment of SLE,systemic sclerosis and vasculitis; KINERET™ (Swedish Oprhan BiovitrumAB), ILARIS™ (Novartis) and ARCALYST™ (Regeneron) which areinterleukin-1 blockers for the treatment of RA and cryopyrin-associatedperiodic syndrome (CAPS).

Cytokines

In some cases, an immunomodulatory composition of the present disclosurecomprises, in addition to CC, a cytokine. Cytokines that can be includedin an immunomodulatory composition of the present disclosure include,without limitation, interleukins, transforming growth factors (TGFs),fibroblast growth factors (FGFs), platelet derived growth factors(PDGFs), epidermal growth factors (EGFs), colony stimulating factors(CSFs), connective tissue activated peptides (CTAPs), osteogenicfactors, and biologically active analogs, fragments, and derivatives ofsuch growth factors. Suitable cytokines include B/T-cell differentiationfactors, B/T-cell growth factors, mitogenic cytokines, chemotacticcytokines, colony stimulating factors, angiogenesis factors, IFN-α,IFN-β, IFN-γ, IL1, IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL9, IL10, IL11,IL12, IL13, IL14, IL15, IL16, IL17, IL18, IL22, etc., leptin, myostatin,macrophage stimulating protein, platelet-derived growth factor, tumornecrosis factor (TNF)-alpha (TNF-α), TNF-β, nerve growth factor (NGF),CD40L, CD137L/4-1BBL, human lymphotoxin-β, G-CSF, M-CSF, GM-CSF,platelet-derived growth factor (PDGF), IL-1α, IL1-β, IP-10, PF4, GRO,9E3, erythropoietin, endostatin, angiostatin, vascular endothelialgrowth factor (VEGF) or any fragments or combinations thereof. Othercytokines include members of the transforming growth factor (TGF)supergene family include the beta transforming growth factors (forexample TGF-β1, TGF-β2, TGF-β3); bone morphogenetic proteins (forexample, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9);heparin-binding growth factors (for example, fibroblast growth factor(FGF), epidermal growth factor (EGF), platelet-derived growth factor(PDGF), insulin-like growth factor (IGF)); hematopoietic growth factors(Flt3); pituitary growth hormones or derivatives; growth hormones,neuroactive hormones, Inhibins (for example, Inhibin A, Inhibin B);differentiation factors (for example, GDF-1); and Activins (for example,Activin A, Activin B, Activin AB). In some cases, an immunomodulatorycomposition of the present disclosure comprises, in addition to CC, acompound or agent modulating cytokines.

Adjuvants

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more adjuvants.

Exemplary adjuvants include, but are not limited to: (1) oil-in-wateremulsion formulations (with or without other specific immunostimulatingagents such as muramyl peptides (see below) or bacterial cell wallcomponents), such as for example (a) MF59™ (WO 90/14837; Chapter 10 inVaccine design: the subunit and adjuvant approach, eds. Powell & Newman,Plenum Press 1995), containing 5% Squalene, 0.5% Tween 80, and 0.5% Span85 (optionally containing MTP-PE) formulated into submicron particlesusing a microfluidizer, (b) SAF, containing 10% Squalane, 0.4% Tween 80,5% pluronic-blocked polymer L121, and thr-MDP either microfluidized intoa submicron emulsion or vortexed to generate a larger particle sizeemulsion, and (c) RIBI™ adjuvant system (RAS), (Ribi Immunochem,Hamilton, Mont.) containing 2% Squalene, 0.2% Tween 80, and one or morebacterial cell wall components such as monophosphorylipid A (MPL),trehalose dimycolate (TDM), and cell wall skeleton (CWS), e.g., MPL+CWS(Detox™); (2) saponin adjuvants, such as QS21 or Stimulon™ (CambridgeBioscience, Worcester, Mass.) may be used or particles generatedtherefrom such as ISCOMs (immunostimulating complexes), which ISCOMS maybe devoid of additional detergent e.g. WO 00/07621; (3) CompleteFreund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA); (4)cytokines, such as interleukins (e.g. IL-1, IL-2, IL-4, IL-5, IL-6,IL-7, IL-12, IL-15, IL-28, etc.) (WO99/44636), etc.), interferons (e.g.gamma interferon), macrophage colony stimulating factor (M-CSF), tumornecrosis factor (TNF), colony-stimulating factors (e.g., GM-CSF), etc.;(5) monophosphoryl lipid A (MPL) or 3-O-deacylated MPL (3dMPL) e.g.GB-2220221, EP-A-0689454, optionally in the substantial absence of alumwhen used with pneumococcal saccharides e.g. WO 00/56358; (6)combinations of 3dMPL with, for example, QS21 and/or oil-in-wateremulsions e.g. EP-A-0835318, EP-A-0735898, EP-A-0761231; (7)oligonucleotides comprising CpG motifs (Krieg Vaccine 2000, 19, 618-622;WO 96/02555, WO 98/16247, WO 98/18810, WO 98/40100, WO 98/55495, WO98/37919 and WO 98/52581), i.e., oligonucleotides containing at leastone CG dinucleotide, where the cytosine is unmethylated; (8) apolyoxyethylene ether or a polyoxyethylene ester e.g. WO 99/52549; (9) apolyoxyethylene sorbitan ester surfactant in combination with anoctoxynol (WO 01/21207) or a polyoxyethylene alkyl ether or estersurfactant in combination with at least one additional non-ionicsurfactant such as an octoxynol (WO 01/21152); (10) a saponin and animmunostimulatory oligonucleotide (e.g. a CpG oligonucleotide) (WO00/62800); (11) an immunostimulant and a particle of metal salt e.g. WO00/23105; (12) a saponin and an oil-in-water emulsion e.g. WO 99/11241;(13) a saponin (e.g. QS21)+3dMPL+IM2 (optionally including a sterol)e.g. WO 98/57659; (14) alphaGalCer and its derivatives; (16) toll-likereceptor (TLR) agonists, NOD-like receptor (NLR) agonists, RIG-Iagonists, agonists for C-type lectin receptors and other pathogenrecognition receptor (PRR) agonists e.g., CpG ODNs, ISS-ODNs,rinatolimod, polyl:C and its derivatives, flagellin, ampligen,imidazoquinalines (e.g., imiquimod, resiquimod), muramyl dipeptides;(17) other substances that act as immunostimulating agents to enhancethe efficacy of the composition. Muramyl peptides includeN-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-25acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP),N-acetylmuramyl-L-alanyl-D-isoglutarninyl-L-alanine-2-(1′-2′-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamineMTP-PE), etc. Adjuvants suitable for administration to a human includedin some cases.

Further exemplary adjuvants include, but are not limited to: choleratoxin B subunit, BCG, Pseudomonas aeruginosa exoprotein A, tocopherol,HBV core, E. coli heat labile toxins (such as LT-A, LT-B), Pertussistoxin, Diphtheria toxoid, tetanus toxoid, Cholera toxin derived(CTA1-DD, CT), mutant LT and CT, Aluminium salt-based adjuvants (such asAlum, Aluminum phosphate, Aluminum sulphate, Alhydrogel), Calciumphosphate, kaolin, monophosphoryl lipid A (MPL^(R)) and its derivatives,glucoppyranosyl lipid A, synthetic lipid A, Lipid A mimetics, Vitamin E,Depovax™, Saponins (Quil-A, ASO1, AS02 (squalene+MPL+QS-21)), AS03, ASO4(alum+MPL^(R)), Tomatin, Protolin, RC-529, Pluronic™, Monatides,Matrix-M, OM-174, Lipovac, IC-31, bacterial/mycobacterial peptides (suchas KLK, cationic (poly)peptides, anti-bacterial microbial peptides,defensins, tuftsin, cathelicidin), dipeptides (such as pidotimod),Bestatin, Hepon (tetradecapeptide), SCV-07(gamma-D-glutamyl-L-tryptophan), Thymosin-a, Immunofan, Thymogen,Indolicidin and its derivatives, polyphosphagene and its derivatives,Gellan, nucleotides (mononucleotides, dinucleotides, polynucleotides,cyclic nucleotides), Eurocine etc.

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more mucoadhesives such as sodium alginate,starch, lectins, thiolated polymers, GelVac™, sodiumcarboxymethylcellulose, hydroxylpropyl methylcellulose, carbomers, cetyltrimethyl ammonium bromide.

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more adjuvant formulations such asoil-in-water emulsions, water-in-oil emulsions, nanoemulsions,particulate delivery systems, liposomes, microspheres, biodegradablemicrospheres, patches virosomes, proteoliposomes, proteasomes,Immunostimulatory complexes (ISCOMs, ISCOMATRIX), microparticles,nanoparticles, biodegradable nanoparticles, silicon nanoparticles,polymeric micro/nano particles, polymeric lamellar substrate particles(PLSP), microparticle resins, nanolipogels, synthetic/biodegradable andbiocompatible semisynthetic or natural polymers or dendrimers (such asPLG, PLGA, PLA, polycaprolactone, silicone polymer, polyesters,poly-dimethyl siloxane, sodium polystyrene sulphonate, polystyrenebenzyl trimethyl ammonium chloride, polystyrene divinyl benzene resin,polyphosphazene, poly-[di-(carboxylactophenoxy)phosphazene] (PCPP),poly-(methylmethacrylate), dextran, polyvinylpyrrolidone, hyaluronicacid and derivatives, chitosan and its derivatives, polysaccharides,Delta inulin polysaccharide, glycolipids (synthetic or natural),lipopolysaccharides, polycationic compound(s) (such as Poly-amino acids,poly-(γ-glutamic acid), poly-arginine-HCl, poly-L-lysine, polypeptides,biopolymers), cationic dimethyldioctadecyl ammonium (DDA),alpha-galactosyl ceramide and its derivatives, archaeal lipids andderivatives, lactanes, gallen, glycerolipids, phospholipids, cochleates,etc. or mixtures thereof.

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more adjuvant formulations such asoil-in-water emulsions or water-in-oil emulsions including edible oils(such as olive oil, mustard oil, vegetable oil, soybean oil, mineral oiletc.).

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more surfactants and detergents (e.g.,non-ionic detergents or niosomes) (such as Tween-80, Polysorbate 80,Span 85, Stearyl tyrosine etc.). An immunomodulatory composition of thepresent disclosure can comprise, in addition to CC, an component oradjuvant mentioned above which provides a depot effect.

Probiotic

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more probiotics. “Probiotic” refers to acomposition containing one species (i.e., a single isolate) or acombination of pure bacteria (i.e., co-culture of desired bacteria), andmay also include any additional carriers, excipients, and/or therapeuticagents that can be administered to a mammal for restoring microbiotaand/or providing health benefits. Examples of probiotics include but arenot limited to, Lactobacillus sp., Bifidobacteria sp., Saccharomycesboulardii, Streptococcus sp., Enterococcus faecium, Bacillus coagulans,Faecalibacterium sp., etc.

Prebiotic

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more prebiotic. As used herein the term“prebiotic” refers to nutritional supplements that are not digested bythe mammal that ingests them, but which are a substrate for the growthor activity of the microbiota, particularly the gut microbiota. Manyprebiotics are carbohydrates, e.g. polysaccharides and oligosaccharides,but the definition does not preclude non-carbohydrates. The mostprevalent forms of prebiotics are nutritionally classed as solublefiber. Prebiotics may provide for changes in the composition and/oractivity of the gastrointestinal microbiota. “Prebiotic” also refers tocompositions containing non-viable food components that are specificallymetabolized in the body by indigenous bacteria thought to be of positivevalue such as Bifidobacteria, Lactobacillus, etc. Examples of prebioticsinclude but are not limited to fructose, xylose, soya, glucose, mannoseetc.

Microbiota

The term “microbiota”, “microbiome”, “symbiotic” or “commensal” usedinterchangeably, refers to microbial population (bacteria, viruses,fungi, parasites) in an individual at various places such as gut, skin,saliva, colon, vagina, lungs etc. A dysbalance in microbiota is relatedto the etiology or onset of several autoimmune and inflammatorydiseases. An immunomodulatory composition of the present disclosure cancomprise, in addition to CC, members of microbiota of an individual suchas Bacteroidetes, Proteobacteria, Firmicutes, Verrucomicrobia,Bacteriodales, Enterobacteriales, Clostridium, etc. Other examples ofmembers of microbiota are known, or will be apparent, to those skilledin the art. See, US Patent Application No. 2014/0010844. See, Howarthand Wang, Nutrients. 2013, 5(1):58-81 for a description of the role ofendogenous microbiota, probiotics and their biological products. Thus acomposition of the invention can be used to establish, modulate,regulate or maintain a balanced microbiota. Members of the microbiomecan be of autologous, allogeneic and xenogeneic origin, wild-type,viable, inactivated, heat-killed, mutated, attenuated and/or geneticallyengineered.

Therapeutic Pathogens

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, therapeutic pathogenic bacteria, virus, fungus etc.such as Listeria, Saccharomyces, Escherichia, Salmonella,Staphylococcus, Klebsiella, poxviruses, adenoviruses, oncolytic viruses.Other examples of therapeutic microbial pathogens are known, or will beapparent, to those skilled in the art. See, US Patent Application No.2014/0010844. Therapeutic pathogen can be wild-type, viable,inactivated, heat-killed, mutated, attenuated and/or geneticallyengineered.

Therapeutic Agents

An immunomodulatory composition of the present disclosure can comprise,in addition to CC, one or more therapeutic agents. Examples oftherapeutic agents are known, or will be apparent, to those skilled inthe art. Non-limiting examples of the therapeutic agents are providedherein the “Methods” section, which include anti-inflammatory agents,anti-proliferative agents, immunosuppressive agents, anti-histamines,immunoregulatory agents, immunomodulatory agents, antimetabolic agents,anti-allergic agents, cytotoxic agents, anti-helminth agents,anti-angiogenic agents, antimicrobial agents (such as antiviral agents,antibacterial agents, anti-parasitic agents, antimalarial agents,anti-protozoal agents), therapeutic peptides etc.

Methods

The present disclosure provides methods of modulating an immune responsein an individual, the method comprising administering to the individualan effective amount of an immunomodulatory composition of the presentdisclosure. The present disclosure provides methods of reducing anundesired immune response in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure. The presentdisclosure provides methods of reducing inflammation in an individual,the method comprising administering to the individual an effectiveamount of an immunomodulatory composition of the present disclosure. Thepresent disclosure provides methods of treating an autoimmune disorderin an individual, the method comprising administering to the individualan effective amount of an immunomodulatory composition of the presentdisclosure. The present disclosure provides methods of treating anallergy (allergic disease) in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure. The presentdisclosure provides methods of treating a metabolic disease in anindividual, the method comprising administering to the individual aneffective amount of an immunomodulatory composition of the presentdisclosure. The present disclosure provides methods of treating aneurological disorder in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure. The presentdisclosure provides methods of enhancing the efficacy and/or reducingthe toxicity of therapeutic treatment in an individual, the methodcomprising administering to the individual an effective amount of animmunomodulatory composition of the present disclosure. The presentdisclosure provides methods of treating, restoring or correctingdisease- or medical condition-related to imbalances in the microbiome ofan individual, the method comprising administering to the individual aneffective amount of an immunomodulatory composition of the presentdisclosure. The present disclosure provides methods of treating,restoring or correcting dysbiosis of an individual, the methodcomprising administering to the individual an effective amount of animmunomodulatory composition of the present disclosure.

The present disclosure also provides a method of modulating dendriticcells, the method comprising: a) contacting dendritic cells (DCs)obtained from an individual with a composition comprising: i)Caulobacter crescentus; and/or ii) an antigen; the contacting step is invitro, and modulates antigen presentation of the antigen on the DCs,thereby generating a population of modulated DCs. The population ofmodulated DCs can then be administered to the individual from whom theDCs were obtained.

In some cases, various immune cells can be obtained from lymphoidtissues, peripheral blood, organs and tissues, and/or can bedifferentiated from stem cells obtained from bone marrow or variousorgans.

The present disclosure also provides a method of inducing proliferation,differentiation and/or modulation of stem cells, the method comprisingcontacting stem cells obtained from an individual with a compositioncomprising Caulobacter crescentus. Contacting the stem cells with the CCleads to proliferation, differentiation and/or modulation of the stemcells, thereby generating a population of expanded, differentiatedand/or modulated cells. The population of expanded, differentiatedand/or modulated cells can then be administered to the individual fromwhom the stem cells were obtained.

The present disclosure further provides a method of generatingregulatory lymphocytes such as NK, NKT, γδ T cells, ILCs, T cells, and Bcells, the method comprising: a) contacting lymphocytes (NK, NKT, γδ Tcells, ILCs, T cells, B cells) obtained from an individual with acomposition comprising: i) Caulobacter crescentus; and/or ii) an antigenin the presence or absence of antigen presenting cells. Contacting thelymphocytes with the CC generates regulatory lymphocytes, therebygenerating a population of regulatory lymphocytes. The population ofregulatory lymphocytes can then be administered to the individual fromwhom the lymphocytes were obtained.

Methods of Modulating an Immune Response

The present disclosure provides methods of modulating an immune responsein an individual, the method comprising administering to the individualan effective amount of an immunomodulatory composition of the presentdisclosure. The present disclosure provides methods of reducing anundesired immune response in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure. The presentdisclosure provides methods of reducing inflammation in an individual,the method comprising administering to the individual an effectiveamount of an immunomodulatory composition of the present disclosure. Thepresent disclosure provides methods of treating an autoimmune disorderin an individual, the method comprising administering to the individualan effective amount of an immunomodulatory composition of the presentdisclosure. The present disclosure provides methods of treating anallergy (allergic disease) in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure.

In some cases, the immune response is a humoral immune response. In somecases, the present disclosure provides methods of modulating a humoralimmune response in an individual, the method comprising administering tothe individual an effective amount of an immunomodulatory composition ofthe present disclosure. In some cases, the immunomodulatory compositiondoes not include any additional antigens (other than antigens present onCC). In some cases, the immunomodulatory composition comprises anantigen (e.g., an antigen other than antigens present on CC). Asdescribed above, suitable antigens include autoantigens, and allergens.

In some cases, the immune response is a cellular immune response. Insome cases, the present disclosure provides methods of modulating acellular immune response in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure. In some cases,the immunomodulatory composition does not include any additionalantigens (other than antigens present on CC). In some cases, theimmunomodulatory composition comprises an antigen (e.g., an antigenother than antigens present on CC). As described above, suitableantigens include autoantigens and allergens.

In some cases, the immune response comprises a modulation in the numberof B cells. In some cases, a subject method comprising administering toan individual in need thereof an effective amount of an immunomodulatorycomposition, where an effective amount of an immunomodulatorycomposition is an amount that, when administered to the individual in asingle dose or in multiple doses, is effective to modulate (e.g.,reduce) the number of B cells in an individual. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to modulate (e.g., reduce) thenumber of B cells in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, or more than 10-fold,compared to the number of B cells in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number ofantigen-specific B cells in an individual. For example, in some cases,an effective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number ofantigen-specific B cells in an individual by at least 10%, at least 15%,at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, or more than 10-fold,compared to the number of antigen-specific B cells in the individual inthe absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce thenumber of antigen-specific B cells in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the number of antigen-specific B cells in the individual inthe absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce thenumber of autoantigen-specific B cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the number of autoantigen-specific B cells in the individualin the absence of treatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce)activation of B cells in an individual. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) activation of Bcells in an individual by at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, at least 25-fold, at least 50-fold,at least 100-fold, or more than 100-fold, compared to the activationlevel of B cells in the individual in the absence of treatment with theimmunomodulatory composition. For example, in some cases, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to reduce the activation of B cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the activation level of B cells in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) theamount of antibody specific to a given antigen in the individual. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate(e.g., reduce) the amount of antibody specific to a given antigen in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, or more than 10-fold, compared to the amountof antibody specific to the antigen in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the amount of antibodyspecific to a given antigen in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of antibody specific to the antigen in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce)production of one or more cytokines in the individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce)production of one or more cytokines in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, or morethan 10-fold, compared to the amount of the cytokine in the individualin the absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce theproduction of one or more cytokines in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the production of one or more cytokines in the individual inthe absence of treatment with the immunomodulatory composition. In othercases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to modulate (e.g., reduce)production of GM-CSF in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, or more than 10-fold,compared to the amount of GM-CSF in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the production of GM-CSFin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the amount of GM-CSF inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) production of IL-22 in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, or more than 10-fold, compared to the amount of IL-22 in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of IL-22 in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of IL-22 in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to modulate (e.g., reduce)production of interferon (IFN)-α and/or IFN-β and/or IFN-γ in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, or more than 10-fold, compared to the amountof IFN-α or IFN-β or IFN-γ in the individual in the absence of treatmentwith the immunomodulatory composition. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to reduce the production of interferon(IFN)-α and/or IFN-β and/or IFN-γ in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of interferon (IFN)-α and/or IFN-β and/or IFN-γin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) production of one or more of IL-17A, IL-2,IL-10, IL-6 and/or TNF-α in an individual by at least 10%, at least 15%,at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, or more than 10-fold,compared to the amount of IL-17A, IL-2, IL-10, IL-6, or TNF-α in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of one or more of IL-17A, IL-2, IL-10, IL-6 andTNF-α in an individual by at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, or more than 75%, compared to the amount ofIL-17A, IL-2, IL-10, IL-6 and TNF-α in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the production of IL-6 inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the amount of IL-6 in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the production of IL-1β in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the amount of IL-1β in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to increase the level of TGF-β in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, or more than 10-fold, compared to the amountof TGF-β in the individual in the absence of treatment with theimmunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., increase,reduce or balance) production of one or more cytokines, chemokines orlymphotoxins such as but not limited to GM-CSF, IL-2, IL-22,Interferons, IL-1β, TGF-β, IL-17A, IL-2, IL-10, IL-6, IL-5, IL-13,TNF-α, IL-9, IL-28 KC/IL-8, MIP-1α, LTα4, etc. in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, or more than 10-fold, compared to the amount of cytokines,chemokines or lymphotoxins such as but not limited to GM-CSF, IL-2,IL-22, Interferons, IL-1β, TGF-β, IL-17A, IL-2, IL-10, IL-6, IL-5,IL-13, TNF-α, IL-9, IL-28, KC/IL-8, MIP-1α, LTα4, etc., in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) a Th1response in an individual. For example, in some cases, an effectiveamount of an immunomodulatory composition of the present disclosure isan amount that is effective, when administered in a single dose or inmultiple doses, to modulate (e.g., reduce) a Th1 response in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the level of the Th1 response in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the Th1 response in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the Th1 response in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD4⁺ T cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD4⁺ T cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of CD4+ T cells in the individual in the absenceof treatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific CD4⁺ T cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific CD4⁺ T cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific CD4⁺ T cellsin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of CD4⁺ T cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of CD4⁺ T cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific CD4+ T cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific CD4⁺ T cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD8⁺ T cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of CD8⁺ T cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of CD8⁺ T cells in the individual in the absenceof treatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific CD8⁺ T cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific CD8⁺ T cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific CD8⁺ T cellsin the individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of CD8⁺ T in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of CD8⁺ T in theindividual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific CD8⁺ T in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific CD8⁺ T in the individual in the absence of treatmentwith the immunomodulatory composition. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to reduce the number and/or activity ofautoantigen-specific CD8⁺ T cells in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the number and/or activity of autoantigen-specific CD8⁺ Tcells in the individual in the absence of treatment with theimmunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of cytolytic T cells in an individual. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate(e.g., reduce) the number and/or activity of cytolytic T cells in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number and/or activity of cytolytic Tcells in the individual in the absence of treatment with theimmunomodulatory composition. In some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) the number and/or activity ofantigen-specific cytolytic T cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific cytolytic T cells in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the number and/or activity ofantigen-specific cytolytic T cells in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to reduce the number and/or activityof cytolytic T cells in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, or more than 75%, compared to thenumber and/or activity of cytolytic T cells in the individual in theabsence of treatment with the immunomodulatory composition. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to reduce the number and/oractivity of antigen-specific cytolytic T cells in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity ofantigen-specific cytolytic T cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate the number and/oractivity of one or more of natural killer (NK) cells, NKT cells, γδ Tcells, ILCs, macrophages, and dendritic cells (DCs) in an individual.For example, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate thenumber and/or activity of one or more of NK cells, NKT cells, γδ Tcells, ILCs, macrophages, and DCs in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of one or more of NK cells, NKT cells, γδ Tcells, ILCs, macrophages, and DCs in the individual in the absence oftreatment with the immunomodulatory composition. For example, in somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to modulate the number and/oractivity of one or more of NK cells, NKT cells, γδ T cells, ILCs,macrophages, and DCs in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, or more than 75%, compared to thenumber and/or activity of NK cells, NKT cells, γδ T cells, ILCs,macrophages, and DCs in the individual in the absence of treatment withthe immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to increase, decrease or balancethe number and/or function of regulatory cells in an individual. Tregs(regulatory T cells) are CD4⁺ or CD8⁺, and may also be FoxP3⁺ Tregs mayalso be defined by other markers such as PD-1, CTLA-4 etc. Regulatorycells may also be comprised of other innate cells such as NK, NKT, yS Tcells, ILCs, and DCs, and B lymphocytes. NK and NKT can also be FoxP3⁺and may also be defined by other markers such as PD-1. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate the number ofregulatory cells in an individual by at least 10%, at least 15%, atleast 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, at least 100% (or 2-fold), atleast 2.5-fold, at least 5-fold, at least 10-fold, more than 10-fold, atleast 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, atleast 100-fold, or more than 100-fold, compared number and/or activityof regulatory cells in the individual in the absence of treatment withthe immunomodulatory composition. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to increase the number of regulatory cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number of regulatory cellsin the individual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th17 cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th17 cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of Th17 cells in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific Th17 cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific Th17 cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific Th17 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of Th17 cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of Th17 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific Th17 cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific Th17 cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th22 cells in an individual. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of Th22 cells in an individual by at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber and/or activity of Th22 cells in the individual in the absence oftreatment with the immunomodulatory composition. In some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) the number and/oractivity of antigen-specific Th22 cells in an individual. For example,in some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) thenumber and/or activity of antigen-specific Th22 cells in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of antigen-specific Th22 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of Th22 cells in an individual byat least 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, ormore than 75%, compared to the number and/or activity of Th22 cells inthe individual in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the number and/or activity of antigen-specific Th22 cells inan individual by at least 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the number and/or activity ofantigen-specific Th22 cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate the number and/oractivity of TH9 cells in an individual. “Modulate the number and/oractivity” of TH9 cells, as used herein, refers to increasing,decreasing, or balancing the number and/or activity of TH9 cells. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to modulate thenumber and/or activity of TH9 cells in an individual by at least 10%, atleast 15%, at least 20%, at least 25%, at least 30%, at least 35%, atleast 40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared numberand/or activity of TH9 cells in the individual in the absence oftreatment with the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to modulate (e.g., reduce) innateand/or adaptive (including both cellular and humoral) immune responsesin an individual. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) the number and/or activity of innate and/oradaptive immune cells and/or their effector functions in an individualby at least 10%, at least 15%, at least 20%, at least 25%, at least 30%,at least 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the number and/or activity of one or more of innate oradaptive immune cells and/or their effector functions in the individualin the absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce thenumber and/or activity of innate and/or adaptive immune cells and/ortheir effector functions in an individual by at least 10%, at least 15%,at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, atleast 45%, at least 50%, at least 75%, or more than 75%, compared to thenumber and/or activity of innate and/or adaptive immune cells and/ortheir effector functions in the individual in the absence of treatmentwith the immunomodulatory composition.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to induce and/or augmentapoptosis in innate and/or adaptive immune cells in an individual toprotect from undesirable inflammation. For example, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to induce and/or augment apoptosis in innateand/or adaptive immune cells in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, at least 100% (or2-fold), at least 2.5-fold, at least 5-fold, at least 10-fold, more than10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least50-fold, at least 100-fold, or more than 100-fold, compared to thenumber of one or more of innate or adaptive immune cells in theindividual in the absence of treatment with the immunomodulatorycomposition.

In some cases, an immunomodulatory composition of the present disclosurecomprises CC and an antigen. Where an immunomodulatory composition ofthe present disclosure comprises CC and an antigen, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) an immune responseto the antigen by at least about 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, more than 10-fold, at least 15-fold,at least 20-fold, at least 25-fold, at least 50-fold, at least 100-fold,or more than 100-fold, compared to the immune response to the antigen inthe absence of treatment with the immunomodulatory composition. Forexample, where the antigen is an antigen associated with or derived froman autoantigen or an allergen, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) an immune response to the antigen by at leastabout 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the immune response to the antigen in the absence oftreatment with the immunomodulatory composition. For example, where theimmunomodulatory composition comprises CC, an antigen, an autoantigen oran allergen, alone or in combination with each other, in some cases, aneffective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to reduce the immune response to the antigenin an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared to the immune response tothe antigen in the individual in the absence of treatment with theimmunomodulatory composition.

The immune response can be a humoral immune response, e.g., a B cell orantibody immune response. Thus, e.g., in some cases, where the antigenis an antigen associated with or derived an autoantigen or an allergen,an effective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) a B cell responseto the antigen by at least about 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, more than 10-fold, at least 15-fold,at least 20-fold, at least 25-fold, at least 50-fold, at least 100-fold,or more than 100-fold, compared to the B cell response to the antigen inthe absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce the Bcell response to the antigen in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the B cell response to the antigen in the individual in theabsence of treatment with the immunomodulatory composition. For example,in some cases, where the antigen is an antigen associated with orderived from an autoantigen or an allergen, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to modulate (e.g., reduce) the amount of antibody specific to theantigen by at least about 10%, at least 15%, at least 20%, at least 25%,at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, at least 100% (or 2-fold), at least 2.5-fold, at least5-fold, at least 10-fold, more than 10-fold, at least 15-fold, at least20-fold, at least 25-fold, at least 50-fold, at least 100-fold, or morethan 100-fold, compared to the amount of antibody specific to theantigen in the absence of treatment with the immunomodulatorycomposition. For example, in some cases, an effective amount of animmunomodulatory composition of the present disclosure is an amount thatis effective, when administered in a single dose or in multiple doses,to reduce the amount of antibody specific to the antigen in anindividual by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 75%, or more than 75%, compared to the amount of antibody specificto the antigen in the individual in the absence of treatment with theimmunomodulatory composition.

The immune response can be a cellular immune response, e.g., a T cellimmune response. Thus, e.g., in some cases, where the antigen is anantigen associated with or derived from an autoantigen or an allergen,an effective amount of an immunomodulatory composition of the presentdisclosure is an amount that is effective, when administered in a singledose or in multiple doses, to modulate (e.g., reduce) a T cell responseto the antigen by at least about 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 35%, at least 40%, at least 45%, atleast 50%, at least 75%, at least 100% (or 2-fold), at least 2.5-fold,at least 5-fold, at least 10-fold, more than 10-fold, at least 15-fold,at least 20-fold, at least 25-fold, at least 50-fold, at least 100-fold,or more than 100-fold, compared to the T cell response to the antigen inthe absence of treatment with the immunomodulatory composition. Forexample, in some cases, an effective amount of an immunomodulatorycomposition of the present disclosure is an amount that is effective,when administered in a single dose or in multiple doses, to reduce the Tcell response to the antigen in an individual by at least 10%, at least15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 75%, or more than 75%,compared to the T cell response to the antigen in the individual in theabsence of treatment with the immunomodulatory composition. In somecases, the immune response is a humoral immune response and a cellularimmune response.

In some cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, to normalize the level of one ormore serum markers of pathological immune responses. For example, insome cases, an effective amount of an immunomodulatory composition ofthe present disclosure is an amount that is effective, when administeredin a single dose or in multiple doses, in normalizing the level of oneor more of the serum markers (these markers include, but are notrestricted to, cholesterol (CHOL), glucose (GLU), globulin (GLOB),alanine aminotransferases (ALT), aspartate aminotransferases (AST),total phosphates (TP), total bilirubin (TBIL), phosphate (PHOS),triglycerides (TRIG), uric acid (URIC), creatine kinase (CK) and urea)in an individual by at least 10%, at least 15%, at least 20%, at least25%, at least 30%, at least 35%, at least 40%, at least 45%, at least50%, at least 75%, or more than 75%, compared with the level of CHOL,GLU, GLOB, ALT, AST, TP, PHOS, TRIG, URIC, CK, TBIL or urea in theindividual in the absence of treatment with the immunomodulatorycomposition.

Adjuvants

In some embodiments, a subject method involves administration of asubject immunomodulatory composition, where the immunomodulatorycomposition comprises CC and one or more adjuvants.

Exemplary adjuvants include, but are not limited to: (1) oil-in-wateremulsion formulations (with or without other specific immunostimulatingagents such as muramyl peptides (see below) or bacterial cell wallcomponents), such as for example (a) MF59^(Tm) (WO 90/14837; Chapter 10in Vaccine design: the subunit and adjuvant approach, eds. Powell &Newman, Plenum Press 1995), containing 5% Squalene, 0.5% Tween 80, and0.5% Span 85 (optionally containing MTP-PE) formulated into submicronparticles using a microfluidizer, (b) SAF, containing 10% Squalane, 0.4%Tween 80, 5% pluronic-blocked polymer L121, and thr-MDP eithermicrofluidized into a submicron emulsion or vortexed to generate alarger particle size emulsion, and (c) RIBI™ adjuvant system (RAS),(Ribi Immunochem, Hamilton, Mont.) containing 2% Squalene, 0.2% Tween80, and one or more bacterial cell wall components such asmonophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wallskeleton (CWS), e.g., MPL+CWS (Detox™); (2) saponin adjuvants, such asQS21 or Stimulon™ (Cambridge Bioscience, Worcester, Mass.) may be usedor particles generated therefrom such as ISCOMs (immunostimulatingcomplexes), which ISCOMS may be devoid of additional detergent e.g. WO00/07621; (3) Complete Freund's Adjuvant (CFA) and Incomplete Freund'sAdjuvant (IFA); (4) cytokines, such as interleukins (e.g. IL-1, IL-2,IL-4, IL-5, IL-6, IL-7, IL-12, IL-15, IL-28, etc.) (WO99/44636), etc.),interferons (e.g. gamma interferon), macrophage colony stimulatingfactor (M-CSF), tumor necrosis factor (TNF), colony-stimulating factors(e.g., GM-CSF), etc.; (5) monophosphoryl lipid A (MPL) or 3-O-deacylatedMPL (3dMPL) e.g. GB-2220221, EP-A-0689454, optionally in the substantialabsence of alum when used with pneumococcal saccharides e.g. WO00/56358; (6) combinations of 3dMPL with, for example, QS21 and/oroil-in-water emulsions e.g. EP-A-0835318, EP-A-0735898, EP-A-0761231;(7) oligonucleotides comprising CpG motifs (Krieg Vaccine 2000, 19,618-622; WO 96/02555, WO 98/16247, WO 98/18810, WO 98/40100, WO98/55495, WO 98/37919 and WO 98/52581), i.e., oligonucleotidescontaining at least one CG dinucleotide, where the cytosine isunmethylated; (8) a polyoxyethylene ether or a polyoxyethylene estere.g. WO 99/52549; (9) a polyoxyethylene sorbitan ester surfactant incombination with an octoxynol (WO 01/21207) or a polyoxyethylene alkylether or ester surfactant in combination with at least one non-ionicsurfactant such as an octoxynol (WO 01/21152); (10) a saponin and animmunostimulatory oligonucleotide (e.g. a CpG oligonucleotide) (WO00/62800); (11) an immunostimulant and a particle of metal salt e.g. WO00/23105; (12) a saponin and an oil-in-water emulsion e.g. WO 99/11241;(13) a saponin (e.g. QS21)+3dMPL+IM2 (optionally including a sterol)e.g. WO 98/57659; (14) alphaGalCer and its derivatives; (16) toll-likereceptor (TLR) agonists, NOD-like receptor (NLR) agonists, RIG-Iagonists, agonists for C-type lectin receptors and other pathogenrecognition receptor (PRR) agonists e.g., CpG ODNs, ISS-ODNs,rinatolimod, polyl:C and its derivatives, flagellin, ampligen,imidazoquinalines (e.g., imiquimod, resiquimod), muramyl dipeptides;(17) other substances that act as immunostimulating agents to enhancethe efficacy of the composition. Muramyl peptides includeN-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-25acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP),N-acetylmuramyl-L-alanyl-D-isoglutarninyl-L-alanine-2-(1′-2′-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamineMTP-PE), etc. Adjuvants suitable for administration to a human includedin some cases.

Further exemplary adjuvants include, but are not limited to: choleratoxin B subunit, BCG, Pseudomonas aeruginosa exoprotein A, tocopherol,HBV core, E. coli heat labile toxins (such as LT-A, LT-B), Pertussistoxin, Diphtheria toxoid, tetanus toxoid, Cholera toxin derived(CTA1-DD, CT), mutant LT and CT, Aluminium salt-based adjuvants (such asAlum, Aluminum phosphate, Aluminum sulphate, Alhydrogel), Calciumphosphate, kaolin, monophosphoryl lipid A (MPL^(R)) and its derivatives,glucoppyranosyl lipid A, synthetic lipid A, Lipid A mimetics, Vitamin E,Depovax™, Saponins (Quil-A, ASO1, AS02 (squalene+MPL+QS-21)), AS03, ASO4(alum+MPL^(R)), Tomatin, Protolin, RC-529, Pluronic™, Monatides,Matrix-M, OM-174, Lipovac, IC-31, bacterial/mycobacterial peptides (suchas KLK, cationic (poly)peptides, anti-bacterial microbial peptides,defensins, tuftsin, cathelicidin), dipeptides (such as pidotimod),Bestatin, Hepon (tetradecapeptide), SCV-07(gamma-D-glutamyl-L-tryptophan), Thymosin-α, Immunofan, Thymogen,Indolicidin and its derivatives, polyphosphagene and its derivatives,Gellan, nucleotides (mononucleotides, dinucleotides, polynucleotides,cyclic nucleotides), Eurocine etc.

Combination Therapy

In some embodiments, a subject method involves administration of asubject immunomodulatory composition as monotherapy, e.g.,administration of a subject immunomodulatory composition only, withoutco-administration of any other therapeutic agent. In other embodiments,a subject treatment method is a combination therapy involvingadministration of: a) a subject immunomodulatory composition; and b) atleast one additional therapeutic agent (or a pharmaceutically acceptablesalt, prodrugs, salts of prodrugs, stereoisomers, tautomers etc. of thetherapeutic agent), where the immunomodulatory composition and the atleast one additional therapeutic agent are administered in combinedamounts that are effective to modulate an immune response. Suitableadditional therapeutic agents are described below.

A subject combination therapy can involve: a) administration of animmunomodulatory composition and at least one additional therapeuticagent at the same time, in the same formulation or in separateformulations; b) administration of at least one additional therapeuticagent within about 5 minutes to about 4 weeks of administration of animmunomodulatory composition, e.g., administration of at least oneadditional therapeutic agent within about 5 minutes to about 15 minutes,within about 15 minutes to about 30 minutes, within about 30 minutes toabout 60 minutes, within about 1 hour to about 2 hours, within about 2hours to about 4 hours, within about 4 hours to about 8 hours, withinabout 8 hours to about 12 hours, within about 12 hours to about 24hours, within about 24 hours to about 2 days, within about 2 days toabout 4 days, within about 4 days to about 7 days, within about 1 weekto about 2 weeks, or within about 2 weeks to about 4 weeks ofadministration of an immunomodulatory composition.

In some embodiments, the at least one additional therapeutic agent isco-formulated with the immunomodulatory composition. In otherembodiments, the at least one additional therapeutic agent and theimmunomodulatory composition are separately formulated.

In some embodiments, an effective amount of an immunomodulatorycomposition and an at least one additional therapeutic agent aresynergistic amounts. As used herein, a “synergistic combination” or a“synergistic amount” of a subject immunomodulatory composition and anadditional (e.g., a second) therapeutic agent is a combination or amountthat is more effective in the therapeutic or prophylactic treatment of adisease than the incremental improvement in treatment outcome that couldbe predicted or expected from a merely additive combination of (i) thetherapeutic or prophylactic benefit of the immunomodulatory compositionwhen administered at that same dosage as a monotherapy and (ii) thetherapeutic or prophylactic benefit of the additional therapeutic agentwhen administered at the same dosage as a monotherapy.

A subject combination therapy can involve: administration of animmunomodulatory composition and at least one additional form of therapysuch as radiation therapy (comprising radioisotopes such as¹²⁵1,strontium-89, ³²P, alpha-emitting isotopes, beta-emitting isotopesetc.), photodynamic therapy, laser therapy, natural product therapy,nutraceutical therapy, cellular therapy, prebiotic therapy, probiotictherapy, symbiotic therapy, paraprobiotic therapy etc., given at thesame or different times.

A subject combination therapy can involve: administration of animmunomodulatory composition and at least one additional form of therapysuch as one or more members of microbiota of an individual such asBacteroidetes, Proteobacteria, Firmicutes, Verrucomicrobia,Bacteriodales, Enterobacteriales, Clostridium, VSL #3 etc. given at thesame or different times. Thus, the present invention can be used toestablish, modulate, regulate or maintain a balanced microbiota. Membersof the microbiome can be wild-type, viable, inactivated, heat-killed,mutated, attenuated and/or genetically engineered.

A subject combination therapy can involve: administration of animmunomodulatory composition and at least one additional form of therapysuch as one or more members of probiotics.

A subject combination therapy can involve: administration of animmunomodulatory composition and at least one additional form of therapysuch as one or more members of therapeutic pathogenic bacteria, virus,fungus etc. such as Listeria, Saccharomyces, Escherichia, Salmonella,Staphylococcus, Klebsiella, poxviruses, adenoviruses, oncolytic viruses.Therapeutic pathogen can be wild-type, mutated, attenuated and/orgenetically engineered. Members of the therapeutic pathogens can bewild-type, viable, inactivated, heat-killed, mutated, attenuated and/orgenetically engineered.

In some embodiments, an effective amount of an immunomodulatorycomposition can be administered in a heterologous or homologousprime-boost vaccine, immunotherapy and/or chemotherapy regimen(s).

A subject combination therapy can involve: administration of animmunomodulatory composition and a therapeutic vaccine.

A subject combination therapy can involve: administration of animmunomodulatory composition and a therapeutic antibody. For example, insome embodiments, a subject method involves: a) administration of animmunomodulatory composition of the present disclosure; and b)administration of at least one antibody. The CC and the antibody can bein the same formulation or in separate formulations. The CC and theantibody can be administered simultaneously, or at different times.Suitable antibodies include an antibody against a cancer antigen or apathogenic antigen (e.g., a therapeutic antibody, monoclonal antibodies,bispecific antibodies, chemoimmuno conjugated antibodies,radioimmunoconjugated antibodies, antibody-cytokine fusion proteins,antibody-antigen fusion proteins, antibody-immunotoxin fusion proteinetc.). Suitable antibodies include, without limitation, antibodiesdirected against co-stimulatory or co-inhibitory molecules (CD28, CD40,ICOS, CD137, OX40, CD137, CD227, CTLA-4, PD-1, KIRs, TCR, PDL1, LAG3,TIM3, VISTA etc.); and other therapeutic antibodies. Non-limitingexamples of suitable antibodies include, but are not limited to,adalimumab, bevacizumab, infliximab, abciximab, alemtuzumab,bapineuzumab, basiliximab, belimumab, briakinumab, brodalumab,canakinumab, certolizumab pegol, cetuximab, conatumumab, denosumab,eculizumab, etrolizumab, gemtuzumab ozogamicin, golimumab, ibritumomabtiuxetan, labetuzumab, mapatumumab, matuzumab, mepolizumab, motavizumab,muromonab-CD3, natalizumab, nimotuzumab, ofatumumab, omalizumab,oregovomab, palivizumab, panitumumab, pemtumornab, pertuzumab,ranibizumab, rituximab, rovelizumab, tocilizumab, tositumomab,trastuzumab, ustekinumab, vedolizomab, zalutumumab, and zanolimumab.

Non-limiting examples of therapeutic and prophylactic antibodies thatcan be used in combination therapy with an immunomodulatory compositionof the present disclosure include MDX-010 (Medarex, N.J.) which is ahumanized anti-CTLA-4 antibody for the treatment of prostate cancer;SYNAGIS™ (MedImmune, Md.) which is a humanized anti-respiratorysyncytial virus (RSV) monoclonal antibody for the treatment of RSVinfection; and HERCEPTIN™ (Trastuzumab) (Genentech, Calif.) which is ahumanized anti-HER2 monoclonal antibody for the treatment of metastaticbreast cancer. Other examples are humanized anti-CD18 F(ab′)₂(Genentech); CDP860 which is a humanized anti-CD18 F(ab′)₂ (Celltech,UK); PRO542 which is an anti-HIV gp120 antibody fused with CD4(Progenics/Genzyme Transgenics); Ostavir which is a human anti-HepatitisB virus antibody (Protein Design Lab/Novartis); PROTOVIR™ which is ahumanized anti-CMV IgGI antibody (Protein Design Lab/Novartis); MAK-195(SEGARD) which is a murine anti-TNF-α F(ab′)₂ (Knoll Pharma/BASF); IC14which is an anti-CD14 antibody (ICOS Pharm); a humanized anti-VEGF IgG1antibody (Genentech); OVAREX™ which is a murine anti-CA 125 antibody(Altarex); PANOREX™ which is a murine anti-17-IA cell surface antigenIgG2a antibody (Glaxo Wellcome/Centocor); BEC2 which is a murineanti-idiotype (GD3 epitope) IgG antibody (ImClone System); IMC-C225which is a chimeric anti-EGFR IgG antibody (ImClone System); VITAXIN™which is a humanized anti-αVβ3 integrin antibody (Applied MolecularEvolution/Medlmmune); Campath 1H/LDP-03 which is a humanized anti-CD52IgG1 antibody (Leukosite); Smart M195 which is a humanized anti-CD33 IgGantibody (Protein Design Lab/Kanebo); RITUXAN™ which is a chimericanti-CD20 IgG1 antibody (IDEC Pharm/Genentech, Roche/Zettyaku);LYMPHOCIDE™ which is a humanized anti-CD22 IgG antibody (Immunomedics);Smart ID10 which is a humanized anti-HLA antibody (Protein Design Lab);ONCOLYM™ (Lym-1) is a radiolabelled murine anti-HLA DIAGNOSTIC REAGENTantibody (Techniclone); ABX-IL8 is a human anti-IL8 antibody (Abgenix);anti-CD11a is a humanized IgG1 antibody (Genentech/Xoma); ICM3 is ahumanized anti-ICAM3 antibody (ICOS Pharm); IDEC-114 is a primatizedanti-CD80 antibody (IDEC Pharm/Mitsubishi); ZEVALIN™ is a radiolabelledmurine anti-CD20 antibody (IDEC/Schering AG); IDEC-131 is a humanizedanti-CD40L antibody (IDEC/Eisai); IDEC-151 is a primatized anti-CD4antibody (IDEC); IDEC-152 is a primatized anti-CD23 antibody(IDEC/Seikagaku); SMART anti-CD3 is a humanized anti-CD3 IgG (ProteinDesign Lab); 5G1.1 is a humanized anti-complement factor 5 (C5) antibody(Alexion Pharm); D2E7 is a humanized anti-TNF-α antibody (CAT/BASF);CDP870 is a humanized anti-TNF-α Fab fragment (Celltech); IDEC-151 is aprimatized anti-CD4 IgG1 antibody (IDEC Pharm/SmithKline Beecham);MDX-CD4 is a human anti-CD4 IgG antibody (Medarex/Eisai/Genmab); CDP571is a humanized anti-TNF-α IgG4 antibody (Celltech); LDP-02 is ahumanized anti-α4β7 antibody (LeukoSite/Genentech); OrthoClone OKT4A isa humanized anti-CD4 IgG antibody (Ortho Biotech); ANTOVA™ is ahumanized anti-CD40L IgG antibody (Biogen); ANTEGREN™ is a humanizedanti-VLA-4 IgG antibody (Elan); MDX-33 is a human anti-CD64 (FcγR)antibody (Medarex/Centeon); SCH55700 is a humanized anti-IL-5 IgG4antibody (Celltech/Schering); SB-240563 and SB-240683 are humanizedanti-IL-5 and IL-4 antibodies, respectively, (SmithKline Beecham);rhuMab-E25 is a humanized anti-IgE IgG1 antibody(Genentech/Norvartis/Tanox Biosystems); ABX-CBL is a murine anti CD-147IgM antibody (Abgenix); BTI-322 is a rat anti-CD2 IgG antibody(Medlmmune/Bio Transplant); Orthoclone/OKT3 is a murine anti-CD3 IgG2aantibody (ortho Biotech); SIMULECT™ is a chimeric anti-CD25 IgG1antibody (Novartis Pharm); LDP-01 is a humanized anti-β₂-integrin IgGantibody (LeukoSite); Anti-LFA-1 is a murine anti CD18 F(ab′).sub.2(Pasteur-Merieux/Immunotech); CAT-152 is a human anti-TGF-β₂ antibody(Cambridge Ab Tech); and Corsevin M is a chimeric anti-Factor VIIantibody (Centocor). The above-listed immunoreactive reagents, as wellas any other immunoreactive reagents, may be administered according toany regimen known to those of skill in the art, including the regimensrecommended by the suppliers of the immunoreactive reagents.

Other examples of therapeutic and prophylactic antibodies that can beused in combination with an immunomodulatory composition of the presentdisclosure include Humira and Remicade; ACTEMRA™ (Genentech) which is arecombinant monoclonal IgG1 anti-human interleukin 6-receptor antibodyfor the treatment of anti-TNF resistant RA and juvenile idiopathicarthritis (JIA); ARZERRA™ (GlaxoSmithKline/Novartis) which is a chimerichuman monoclonal antibody directed against membrane proximal epitope onthe CD20 molecule for the treatment of RA; BENLYSTA™ (GlaxoSmithKline)which is a human monoclonal IgG1 gamma that binds to and inhibits thesoluble form of the B-lymphocyte stimulator (BLyS) protein for thetreatment of SLE; ORENCIA™ (Bristol-Myers Squibb) which is a CTLA-4 IgG1binding to CD80/86 on antigen-presenting cells inhibiting theco-stimulation of CD28 on the T cells for the treatment of RA, JIA andSLE; SIMPONI (Janssen) which is a IgG1 monoclonal antibody acting onboth soluble and membrane-bound TNF-α for the treatment of RA, psoriaticarthritis (PsA) and ankylosing spondylitis (AS); CIMZIA™ (UCB Group)which is a pegylated humanized antibody Fab′ fragment of the TNF-αmonoclonal antibody for the treatment of RA; Sifalimumab (MedImmune)which is an anti-IFN-α monoclonal antibody designed for the treatment ofSLE, dematomyositis and polymyositis; various intravenous immunoglobulinproducts which are pools of immunoglobulins from healthy individuals forthe treatment of SLE, systemic sclerosis and vasculitis; KINERET™(Swedish Oprhan Biovitrum AB), ILARIS™ (Novartis) and ARCALYST™(Regeneron) which are interleukin-1 blockers for the treatment of RA andcryopyrin-associated periodic syndrome (CAPS).

A subject combination therapy can involve: administration of animmunomodulatory composition of the present disclosure and one or morecytokines. For example, in some embodiments, a subject method involves:a) administration of an immunomodulatory composition of the presentdisclosure; and b) administration of one or more cytokines. The CC andthe one or more cytokines can be in the same formulation or in separateformulations. The CC and the one or more cytokines can be administeredsimultaneously, or at different times. Suitable cytokines include,without limitation, interleukins, transforming growth factors (TGFs),fibroblast growth factors (FGFs), platelet derived growth factors(PDGFs), epidermal growth factors (EGFs), colony stimulating factors(CSFs), connective tissue activated peptides (CTAPs), osteogenicfactors, and biologically active analogs, fragments, and derivatives ofsuch growth factors. Suitable cytokines include B/T-cell differentiationfactors, B/T-cell growth factors, mitogenic cytokines, chemotacticcytokines, colony stimulating factors, angiogenesis factors, IFN-α,IFN-β, IFN-γ, IL1, IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL9, IL10, IL11,IL12, IL13, IL14, IL15, IL16, IL17, IL18, IL22, etc., leptin, myostatin,macrophage stimulating protein, platelet-derived growth factor, tumornecrosis factor (TNF)-alpha (TNF-α), TNF-β, nerve growth factor (NGF),CD40L, CD137L/4-1BBL, human lymphotoxin-β, G-CSF, M-CSF, GM-CSF,platelet-derived growth factor (PDGF), IL-1α, IL1-β, IP-10, PF4, GRO,9E3, erythropoietin, endostatin, angiostatin, vascular endothelialgrowth factor (VEGF) or any fragments or combinations thereof. Othercytokines include members of the transforming growth factor (TGF)supergene family include the beta transforming growth factors (forexample TGF-β1, TGF-β2, TGF-(33); bone morphogenetic proteins (forexample, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9);heparin-binding growth factors (for example, fibroblast growth factor(FGF), epidermal growth factor (EGF), platelet-derived growth factor(PDGF), insulin-like growth factor (IGF)); hematopoietic growth factors(Flt3); pituitary growth hormones or derivatives; growth hormones,neuroactive hormones, Inhibins (for example, Inhibin A, Inhibin B);differentiation factors (for example, GDF-1); and Activins (for example,Activin A, Activin B, Activin AB).

A subject combination therapy can involve: administration of animmunomodulatory composition of the present disclosure and one or moretherapeutic agents such as anti-angiogenic agents (e.g., in methods forthe treatment of solid tumors and for the treatment and prevention ofmetastases) and anti-hormonal agents (particularly in methods for thetreatment of hormone-dependent cancers such as breast cancer andprostate cancer).

In one embodiment, an immunomodulatory composition of the presentdisclosure is administered in combination with one or moreanti-angiogenic agents. Such agents include, without limitation,angiostatin, thalidomide, kringle 5, endostatin, Serpin (Serine ProteaseInhibitor) anti-thrombin, 29 kDa N-terminal and a 40 kDa C-terminalproteolytic fragments of fibronectin, 16 kDa proteolytic fragment ofprolactin, 7.8 kDa proteolytic fragment of platelet factor-4, a 13-aminoacid peptide corresponding to a fragment of platelet factor-4 (Maione etal., 1990, Cancer Res. 51:2077-2083), a 14-amino acid peptidecorresponding to a fragment of collagen I (Tolma et al., 1993, J. CellBiol. 122:497-511), a 19 amino acid peptide corresponding to a fragmentof Thrombospondin I (Tolsma et al., 1993, J. Cell Biol. 122:497-511), a20-amino acid peptide corresponding to a fragment of SPARC (Sage et al.,1995, J. Cell. Biochem. 57:1329-1334), or any fragments, family members,or variants thereof, including pharmaceutically acceptable saltsthereof.

Other peptides that inhibit angiogenesis and correspond to fragments oflaminin, fibronectin, procollagen, and EGF have also been described(see, e.g., Cao, 1998, Prog Mol Subcell Biol. 20:161-176). Monoclonalantibodies and cyclic pentapeptides, which block certain integrins thatbind RGD proteins (i.e., possess the peptide motif Arg-Gly-Asp), havebeen demonstrated to have anti-vascularization activities (Brooks etal., 1994, Science 264:569-571; Hammes et al., 1996, Nature Medicine2:529-533). Moreover, inhibition of the urokinase plasminogen activatorreceptor by receptor antagonists inhibits angiogenesis, tumor growth andmetastasis (Min et al., 1996, Cancer Res. 56: 2428-33; Crowley et al.,1993, Proc Natl Acad Sci. 90:5021-25).

In another embodiment, a combination therapy of the present disclosurecomprises administering an immunomodulatory composition of the presentdisclosure together with a hormonal treatment modality. Such treatmentmodalities include the administration of hormonal antagonists (e.g.,flutamide, bicalutamide, tamoxifen, raloxifene, leuprolide acetate(LUPRON), LH-RH antagonists), inhibitors of hormone biosynthesis andprocessing, and steroids (e.g., dexamethasone, retinoids, deltoids,betamethasone, cortisol, cortisone, prednisone, dehydrotestosterone,glucocorticoids, mineralocorticoids, estrogen, testosterone,progestins), vitamin A derivatives (e.g., all-trans retinoic acid(ATRA)); vitamin D3 analogs; antigestagens (e.g., mifepristone,onapristone), and antiandrogens (e.g., cyproterone acetate).

In another embodiment, an immunomodulatory composition of the presentdisclosure is used in association with a treatment modality thatutilizes polynucleotide compounds, such as antisense polynucleotides,ribozymes, RNA interference molecules, triple helix polynucleotides andthe like.

In certain embodiments, an immunomodulatory composition of the presentdisclosure is administered in combination with an immunoregulatoryagent. In some embodiments, the immunomodulatory composition isformulated with the immunoregulatory agent. An “immunoregulatory agent”is a substance that suppresses, masks, or enhances the immune system ofthe subject to whom it is administered. Exemplary agents are those thatsuppress cytokine production, downregulate or suppress self-antigenexpression, or mask the MHC antigens. Examples of such agents include2-amino-6-aryl-5-substituted pyrimidines (see, U.S. Pat. No. 4,665,077),azathioprine (or cyclophosphamide, if there is an adverse reaction toazathioprine); bromocryptine; glutaraldehyde (which masks the MHCantigens, as described in U.S. Pat. No. 4,120,649); anti-idiotypicantibodies for MHC antigens and MHC fragments; cyclosporin A; steroidssuch as glucocorticosteroids, e.g., prednisone, methylprednisolone, anddexamethasone; cytokine or cytokine receptor antagonists includinganti-interferon-γ, -β, or a antibodies; anti-tumor necrosis factor-aantibodies; anti-tumor necrosis factor-.beta. antibodies;anti-interleukin-2 antibodies and anti-IL-2 receptor antibodies;anti-L3T4 antibodies; heterologous anti-lymphocyte globulin; pan-Tantibodies, preferably anti-CD3 or anti-CD4/CD4a antibodies; solublepeptide containing a LFA-3 binding domain; IDO inhibitors;streptokinase; TGF-β; streptodomase; FK506; RS-61443; deoxyspergualin;and rapamycin. Examples of cytokines include, but are not limited tolymphokines, monokines, and traditional polypeptide hormones. Includedamong the cytokines are growth hormone such as human growth hormone,N-methionyl human growth hormone, and bovine growth hormone; parathyroidhormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin;glycoprotein hormones such as follicle stimulating hormone (FSH),glucagon, thyroid stimulating hormone (TSH), and luteinizing hormone(LH); hepatic growth factor; fibroblast growth factor; prolactin;placental lactogen; tumor necrosis factor-α; mullerian-inhibitingsubstance; mouse gonadotropin-associated peptide; inhibin; activin;vascular endothelial growth factor; integrin; thrombopoiotin (TPO);nerve growth factors such as NGF-α; platelet-growth factor; transforminggrowth factors (TGFs) such as TGF-α and TGF-β; insulin-like growthfactor-I and —II; erythropoietin (EPO); osteoinductive factors;interferons; colony stimulating factors (CSFs) such as macrophage-CSF(M-CSF); granulocyte-macrophage-CgP (GM-CSP); and granulocyte-CSF(G-CSF); interleukins (ILs) such as IL-1, IL-1α, IL-2, IL-3, IL-4, IL-5,IL-6, IL-7, IL-8, IL-9, IL-11, IL-12, IL-15; a tumor necrosis factorsuch as TNF-α or TNF-β; and other polypeptide factors including LIF andkit ligand (KL). As used herein, the term cytokine includes proteinsfrom natural sources or from recombinant cell culture and biologicallyactive equivalents of the native sequence cytokines. Other examples ofimmunoregulatory agents include mesalazine, mesalamine, sulfasalazine,sulfasalazine derivatives, anti-histamines, glucocorticoids,epinephrine, theophylline, cromolyn sodium, anti-leukotrienes,anti-cholinergic drugs for rhinitis, anti-cholinergic decongestants,mast-cell stabilizers, monoclonal anti-IgE antibodies.

In certain embodiments, an immunomodulatory composition of the presentdisclosure is administered in combination therapy with one or moreimmunomodulatory agents, e.g., a cytokine. Suitable cytokines include,but are not limited to, interleukin-1 (IL-1), IL-2, IL-3, IL-12, IL-15,IL-18, G-CSF, GM-CSF, thrombopoietin, and γ interferon.

Methods of Modulating an Anti-Bacterial Immune Response

The present disclosure provides methods of modulating an immune responseto a bacterium or a substance produced by a bacterium, the methodcomprising administering to an individual in need thereof an effectiveamount of an immunomodulatory composition of the present disclosure.

In some cases, a method of the present disclosure of modulating animmune response to a bacterium, or a substance produced by a bacterium,is effective to reduce the number of bacteria (e.g., pathogenicbacteria) in the individual by at least about 25%, at least about 50%,at least about 75%, or at least about 99%, compared to a pre-treatmentnumber of pathogenic bacteria in the individual, or to an extent thatthe pathogenic bacterium cannot be detected in the individual (e.g., ina biological sample obtained from the individual).

In some cases, a method of the present disclosure of modulating animmune response to a bacterium, or a substance produced by a bacterium(such as endotoxins, toxins, LPS etc.), is effective to regulate animmune response to a pathogenic bacterium. Pathogenic bacteria include,e.g., Gram positive bacteria, Gram negative bacteria, mycobacteria, etc.Non-limiting examples of pathogenic bacteria include Mycobacteria,Streptococcus, Staphylococcus, Pseudomonas, Salmonella, Neisseria, andListeria. In some cases, the bacteria is Neisseria gonorrhea, M.tuberculosis, M. leprae, Listeria monocytogenes, Streptococcuspneumoniae, S. pyogenes, S. agalactiae, S. viridans, S. faecalis, S.aureus, S. epidermis, or S. Bovis.

Other examples of pathogenic bacteria contemplated include, but are notlimited to, Gram positive bacteria (e.g., Listeria, Bacillus such asBacillus anthracis, Erysipelothrix species), Gram negative bacteria(e.g., Bartonella, Brucella, Burkholderia, Campylobacter, Enterobacter,Escherichia, Francisella, Hemophilus, Klebsiella, Morganella, Proteus,Providencia, Pseudomonas, Salmonella, Serratia, Shigella, Vibrio, andYersinia species), spirochete bacteria (e.g., Borrelia species includingBorrelia burgdorferi that causes Lyme disease), anaerobic bacteria(e.g., Actinomyces and Clostridium species), Gram positive and negativecoccal bacteria, Enterococcus species, Streptococcus species,Pneumococcus species, Staphylococcus species, Neisseria species.

Additional non-limiting examples of specific infectious bacteria includeCitrobacter, Chlamydia spp., Helicobacter pyloris, Borelia burgdorferi,Legionella pneumophila, Mycobacteria avium, M. intracellulare, M.kansaii, M. gordonae, M. africanum, Staphylococcus aureus, Neisseriameningitidis, Haemophilus influenzae, Bacillus anthracis, Yersiniapestis, Corynebacterium diphtheriae, Erysipelothrix rhusiopathiae,Clostridium perfringens, Clostridium tetani, Enterobacter aerogenes,Klebsiella pneumoniae, Pasteurella multocida, Fusobacterium nucleatum,Streptobacillus moniliformis, Treponema pallidium, Treponema pertenue,Leptospira, Rickettsia, Porphyromonas gingivalis, and Actinomycesisraelli.

The pathogenic bacteria can be wild-type, viable, inactivated,heat-killed, mutated, attenuated and/or genetically modified.

In some cases, a method of the present disclosure of modulating animmune response to a bacterium, or a substance produced by a bacterium,comprises administering an immunomodulatory composition to an individualin need thereof, and further comprising administering to the individualan effective amount of an anti-bacterial or an antimycobacterial agent.Anti-bacterial and anti-mycobacterial agents are known in the art andinclude, e.g., beta-lactam antibiotics, tetracyclines, streptomycin,chloramphenicol, neomycin, gramicidin, bacitracin, sulfonamides,nitrofurazone, nalidixic acid, rifampicin, fluoroquinolones, isoniazid,pyrazinamide, vancomycin, methicillin etc.

Suitable anti-bacterial agents include, e g, Aminoglycosides such asAmikacin, Apramycin, Arbekacin, Bambermycins, Butirosin, Dibekacin,Dihdrostreptomycin, Fortimicin(s), Gentamicin, Ispamicin, Kanamycin,Micronomicin, Neomycin, Neomycin Undecylenate, Netilmicin, Paromomycin,Ribostamycin, Sisomicin, Spectinomycin, Streptomycin, Streptonicozid andTobramycin; Ansamycins such as Rifamide, Rifampin, Rifamycin andRifaximin; β-lactams such as Carbapenems such as Imipenem;Cephalosporins such as Cefactor, Cefadroxil, Cefamandole, Cefatrizine,Cefazedone, Cefazolin, Cefixime, Cefinenoxime, Cefodizime, Cefonicid,Cefoperazone, Ceforanide, Cefotaxime, Cefotiam, Cefpimizole,Cefpirimide, Cefpodoxime Proxetil, Cefroxadine, Cefsulodin, Ceftazidime,Cefteram, Ceftezole, Ceftibuten, Ceftizoxime, Ceftriaxone, Cefuroxime,Cefuzonam, Cephacetrile Sodium, Cephalexin, Cephaloglycin,Cephaloridine, Cephalosporin, Cephalothin, Cephapirin Sodium, Cephradineand Pivcefalexin; Cephamycins such as Cefbuperazone, Cefmetazole,Cefminox, Cefetan and Cefoxitin; Monobactams such as Aztreonam,Carumonam and Tigemonam; Oxacephems such as Flomoxef and Moxolactam;Penicillins such as Amidinocillin, Amdinocillin Pivoxil, Amoxicillin,Ampicillan, Apalcillin, Aspoxicillin, Azidocillan, Azlocillan,Bacampicillin, Benzylpenicillinic Acid, Benzylpenicillin Sodium,Carbenicillin, Carfecillin Sodium, Carindacillin, Clometocillin,Cloxacillin, Cyclacillin, Dicloxacillin, Diphenicillin Sodium,Epicillin, Fenbenicillin, Floxicillin, Hetacillin, Lenampicillin,Metampicillin, Methicillin Sodium, Mezlocillin, Nafcillin Sodium,Oxacillin, Penamecillin, Penethamate Hydriodide, Penicillin GBenethamine, Penicillin G Benzathine, Penicillin G Benzhydrylamine,Penicillin G Calcium, Penicillin G Hydrabamine, Penicillin G Potassium,Penicillin G Procaine, Penicillen N, Penicillin O, Penicillin V,Penicillin V Benzathine, Penicillin V Hydrabamine, Penimepicycline,Phenethicillin Potassium, Piperacillin, Pivapicillin, Propicillin,Quinacillin, Sulbenicillin, Talampicillin, Temocillin and Ticarcillin;Lincosamides such as Clindamycin and Lincomycin; Macrolides such asAzithromycin, Carbomycin, Clarithromycin, Erythromycin, ErythromycinAcistrate, Erythromycin Estolate, Erythromycin Glucoheptonate,Erythromycin Lactobionate, Erythromycin Propionate, ErythromycinStearate, Josamycin, Leucomycins, Midecamycins, Miokamycin,Oleandomycin, Primycin, Rokitamycin, Rosaramicin, Roxithromycin,Spiramycin and Troleandomycin; Polypeptides such as Amphomycin,Bacitracin, Capreomycin, Colistin, Enduracidin, Enviomycin, Fusafungine,Gramicidin(s), Gramicidin S, Mikamycin, Polymyxin, PolymyxinB-Methanesulfonic Acid, Pristinamycin, Ristocetin, Teicoplanin,Thiostrepton, Tuberactinomycin, Tyrocidine, Tyrothricin, Vancomycin,Viomycin, Viomycin Pantothenate, Virginiamycin and Zinc Bacitracin;Tetracyclines such as Apicycline, Chlortetracycline, Clomocycline,Demeclocycline, Doxycycline, Guamecycline, Lymecycline, Meclocycline,Methacycline, Minocycline, Oxytetracycline, Penimepicycline,Pipacycline, Rolitetracycline, Sancycline, Senociclin and Tetracycline;Cycloserine; Mupirocin; and Tuberin. Suitable anti-bacterial agentsinclude antibodies specific for a bacterium.

Methods of Modulating an Anti-Viral Immune Response

The present disclosure provides methods of modulating an immune responseto a virus, the method comprising administering to an individual in needthereof an effective amount of an immunomodulatory composition of thepresent disclosure.

In some cases, a method of the present disclosure of modulating animmune response to a virus is effective to reduce the number of viruses(e.g., pathogenic viruses) in the individual by at least about 25%, atleast about 50%, at least about 75%, or at least about 99%, or to anextent that the pathogenic virus cannot be detected in the individual(e.g., in a biological sample obtained from the individual).

For example, in some cases, a method of the present disclosure ofmodulating an immune response to a virus is effective to reduce theviral load in the individual by at least about 25%, at least about 50%,at least about 75%, or at least about 99%, or to an extent that thepathogenic virus cannot be detected in the individual (e.g., in abiological sample obtained from the individual). In some cases, a methodof the present disclosure of modulating an immune response to a virus iseffective to reduce the number of genome copies of the virus in theindividual by at least about 25%, at least about 50%, at least about75%, or at least about 99%, compared to a pre-treatment number of genomecopies of the virus in the individual, or to an extent that no genomecopies of the virus can be detected in the individual (e.g., in abiological sample obtained from the individual).

In some cases, a method of the present disclosure of modulating animmune response to a virus regulates an immune response to a pathogenicvirus. Pathogenic viruses include, but are not limited to, herpesviruses (HSV-1, HSV-2, VZV, EBV, CMV, HHV-6, HHV-8), influenza viruses(Flu A, B), hepatitis viruses (HepA, HepB, HepC, HepD, HepE), humanimmunodeficiency viruses (HIV-1, HIV-2), respiratory syncytial viruses,measles viruses, rhinoviruses, adenoviruses, SARS viruses,papillomaviruses, orthopoxviruses, West Nile viruses, and a dengueviruses. Pathogenic viruses include members of the Flaviviridae familyof viruses. Pathogenic viruses include a flavivirus selected from thegroup consisting of dengue, Kunjin, Japanese encephalitits, West Nile,and yellow fever virus. Pathogenic viruses include lymphocyticchoriomenignitis virus, hepatitis B virus, Epstein Barr virus, and humanimmunodeficiency virus. Pathogenic viruses include, but are not limitedto: Retroviridae (e.g. human immunodeficiency viruses, such as HIV-1,also referred to as LAV or HTLV-III/LAV, or HIV-III; and other isolates,such as HIV-LP; Picornaviridae (e.g. polio viruses, hepatitis A virus;enteroviruses, human Coxsackie viruses, rhinoviruses, echoviruses);Calciviridae (e.g. strains that cause gastroenteritis); Togaviridae(e.g. equine encephalitis viruses, rubella viruses); Flaviridae (e.g.dengue viruses, encephalitis viruses, yellow fever viruses);Coronaviridae (e.g. coronaviruses); Rhabdoviridae (e.g. vesicularstomatitis viruses, rabies viruses); Filoviridae (e.g. ebola-likeviruses; Marburg virus); Paramyxoviridae (e.g. parainfluenza viruses,mumps virus, measles virus, respiratory syncytial virus);Orthomyxoviridae (e.g. influenza viruses); Bungaviridae (e.g. Hantaanviruses, bunga viruses, phleboviruses and Nairo viruses); Arenaviridae(hemorrhagic fever viruses); Reoviridae (e.g. reoviruses, orbiviursesand rotaviruses); Bornaviridae; Hepadnaviridae (Hepatitis B virus);Parvoviridae (parvoviruses); Papovaviridae (papilloma viruses, polyomaviruses); Adenoviridae (e.g., adenoviruses); Herpesviridae (herpessimplex virus (HSV) 1 and 2), varicella zoster virus, cytomegalovirus(CMV), herpes virus; Poxviridae (variola viruses, vaccinia viruses, poxviruses); and Iridoviridae (e.g. African swine fever virus); andunclassified viruses (e.g. the etiological agents of Spongiformencephalopathies, the agent of delta hepatitis, thought to be adefective satellite of hepatitis B virus), the agents of non-A, non-Bhepatitis (class 1, internally transmitted; class 2, parenterallytransmitted, i.e., Hepatitis C Virus); Norwalk and related viruses, andastroviruses.

In some cases, a method of the present disclosure of modulating animmune response to a virus, comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of at least oneadditional therapeutic agent, e.g., an anti-viral agent.

Anti-viral agents are known in the art and include, e.g., an anti-HCVagent such as ribavirin and its analogues; glycosidase inhibitors;glucosidase inhibitors; IRES (internal ribosomal entry site), p7, entry,fusion, helicase, assembly, egress, NS2, NS3, NS4, NS5a and NS5Binhibitors; inosine monophosphate dehydrogenase inhibitors; cyclophilininhibitors; metalloprotease inhibitors; anti-HCV nucleos(t)ide andnon-nucleoside RNA polymerase inhibitors etc.; an anti-HIV agent;anti-HBV agent; and the like.

In some embodiments, the at least one additional therapeutic agent is aninterferon (e.g., interferon-alpha, interferon-beta, interferon-gamma,interferon-lambda, interferon-tau, interferon-omega, etc.). In someembodiments, the at least one additional therapeutic agent is IFN-α. Insome embodiments, the at least one additional therapeutic agent isIFN-β.

Suitable additional anti-viral agents for treating an HCV infectioninclude, but are not limited to, ribavirin and its prodrugs such asviramidine, telaprevir, sofosbuvir, boceprevir, ciluprevir, simeprevir,danoprevir, vaniprevir, MK-5172, MK-0608, 2′-C-methyl-7-deaza adenosine,2′-C-methyl-adenosines, BI201335, narlaprevir, asunaprevir, GS-9256,GS-9451, ABT-450, IDX-320, ACH-1625, Valopicitabine, mericitabine,R1626, PSI-938, INX-189, BILN1941, BI-207127, VCH222, VX-135, ANA598,ANA773, ABT-072, ABT-333, HCV-796, GS-9190, Daclatasavir, BMS-824393,BMS-791325, PPI-461, GS-5885, alisporivir (Debio-025), NIM-811, SCY-635,nitazoxanide, clemizole, miravirasen, celgosivir, BCX-5191, GSK-2336805,anti-PD-1 antibodies (CT-011), bavituximab (anti-phosphatidyl serineMab), therapeutic vaccine (GI-5005, IC-41, TG-4040) prophylactic vaccine(such as HCV E1/E2/MF-59), and the prodrugs thereof. Suitable additionaltherapeutic agents include, e.g., therapeutic agents for the treatmentof an hepatitis B virus infection include, but are not limited tolamivudine, adefovir, entecavir, telbuvudine, tenofovir and the prodrugsthereof.

For example, suitable additional anti-viral agents for treating an HCVinfection include weekly injections of pegylated IFN-α combined withtwice-daily oral doses of ribavirin(1-β-D-ribofuranosyl-1H-1,2,4-triazole-3-carboxamide).

Suitable additional therapeutic agents include, e.g., therapeutic agentsfor the treatment of an immunodeficiency virus infection, or for thetreatment of a disorder that may accompany an immunodeficiency virusinfection (e.g., a bacterial infection, a fungal infection, and thelike). Suitable additional therapeutic agents include, e.g., beta-lactamantibiotics, tetracyclines, chloramphenicol, neomycin, gramicidin,bacitracin, sulfonamides, nitrofurazone, nalidixic acid, cortisone,hydrocortisone, betamethasone, dexamethasone, fluocortolone,prednisolone, triamcinolone, indomethacin, sulindac, acyclovir,amantadine, rimantadine, recombinant soluble CD4 (rsCD4), cyanovirin-N,microvirin, fuzeon, anti-receptor antibodies (e.g., for rhinoviruses),nevirapine, cidofovir (Vistide™), trisodium phosphonoformate(Foscarnet™), famcyclovir, pencyclovir, valacyclovir, nucleicacid/replication inhibitors, interferon, zidovudine (AZT, Retrovir™),didanosine (dideoxyinosine, ddl, Videx™), stavudine (d4T, Zerit™)zalcitabine (dideoxycytosine, ddC, Hivid™), nevirapine (Viramune™),lamivudine (Epivir™, 3TC), protease inhibitors, saquinavir (Invirase™,Fortovase™), ritonavir (Norvir™), nelfinavir (Viracept™), efavirenz(Sustiva™), abacavir (Ziagen™) amprenavir (Agenerase™) indinavir(Crixivan™), ganciclovir, AzDU, delavirdine (Rescriptor™), kaletra,trizivir, rifampin, clathiromycin, erythropoietin, colony stimulatingfactors (G-CSF and GM-CSF), non-nucleoside reverse transcriptaseinhibitors, nucleoside inhibitors, viral entry inhibitors, fusioninhibitors, integrase inhibitors, adriamycin, fluorouracil,methotrexate, asparaginase and combinations thereof. Additional suitabletherapeutic agents for HIV include integrase and fusion inhibitors suchas Raltegravir, Elvitegravir, Enfuvirtide, Maraviroc etc.

In some embodiments, the at least one additional therapeutic agent is aneuraminidase inhibitor, e.g., where the influenza virus is influenza Aor influenza B. Suitable neuraminidase inhibitors include, e.g.,oseltamivir (ethyl(3R,4R,5S)-5-amino-4-acetamido-3-(pentan-3-yloxy)cyclohex-1-ene-1-carboxylate;Tamiflu™), zanamivir(2R,3R,4S)-4-[(diaminomethylidene)amino]-3-acetamido-2[(1R,2R)-1,2,3-trihydroxypropyl]-3,4-dihydro-2H-pyran-6-carboxylicacid; Relenza™), and peramivir(1S,2S,3S,4R)-3-[(1S)-1-acetamido-2-ethyl-butyl]-4-(diaminomethylideneamino)-2-hydroxy-cyclopentane-1-carboxylicacid). In some embodiments, the at least one additional therapeuticagent is an M2 blocker, e.g., blocks a viral ion channel (M2 protein).The antiviral drugs amantadine and rimantadine are M2 blockers, and canbe used in subject method.

Suitable additional therapeutic agents, e.g., for the treatment of anHSV-1 or an HSV-2 infection include, but are not limited to, acyclovir(Zovirax), valganciclovir, famciclovir, valacyclovir (Valtrex),ganciclovir (Cytovene), cidofovir (Vistide), antisense oligonucleotidefomivirsen (Vitravene), foscarnet (Foscavir), penciclovir, idoxuridine,vidarabine, and trifluridine.

In some embodiments, the one or more different therapeutic agent isselected antiviral agents that target two or more different viruses;e.g., an HIV inhibitor, HBV inhibitor, HCV inhibitor, herpes virusinhibitor, influenza virus inhibitor, RNA inhibitor, interfering RNA(RNAi) inhibitor, natural products etc. In some cases, a method of thepresent disclosure of treating a viral infection comprises administeringan immunomodulatory composition to an individual in need thereof, andfurther comprising administering to the individual an effective amountof at least one additional therapeutic agent, e.g., a monoclonalantibody or antibody products directed against viral antigens, wheresuitable monoclonal antibodies include but are not limited to HBIg,antibodies against influenza virus strains, anti-hepatitis A virusantibody, SYNAGIS (anti-RSV Mab), anti-rabies antibody, ostavir(anti-HBV Mab), Pro542 (anti-HIV gp120), Potovir (anti-CMV Mab),anti-PD-1 antibodies (CT-011), bavituximab (anti-phosphatidyl serineMab) etc.

Methods of Modulating an Immune Response to a Parasitic Infection

The present disclosure provides methods of modulating an immune responseto a microbial parasite (e.g., a pathogenic protozoan; a helminth;etc.), the method comprising administering to an individual in needthereof an effective amount of an immunomodulatory composition of thepresent disclosure.

In some cases, a method of the present disclosure of modulating animmune response to a microbial parasite is effective to reduce thenumber of microbial parasites (e.g., pathogenic protozoa; pathogenichelminths) in the individual by at least about 25%, at least about 50%,at least about 75%, or at least about 99%, compared to a pre-treatmentnumber of microbial parasite in the individual, or to an extent that themicrobial parasite cannot be detected in the individual (e.g., in abiological sample obtained from the individual).

In some cases, a method of the present disclosure of modulating animmune response to a microbial parasite comprises administering animmunomodulatory composition to an individual in need thereof, andfurther comprising administering to the individual an effective amountof a least one additional therapeutic agent. Anti-parasitic agents areknown in the art and include, e.g., chloroquine, etc. For example,anti-malarial agents include, e.g., quinine, chloroquine, atovaquone,proguanil, primaquine, amodiaquine, mefloquine, piperaquine,artemisinin, methylene blue, pyrimethamine, sulfadoxine,artemether-lumefantrine, dapsone-chlorproguanil, artesunate, quinidine,clopidol, pyridine/pyridinol analogs, 4(1H)-quinolone analogs,dihydroartemisinin, a mixture of atovaquone and proguanil, anendoperoxide, and an acridone. Anti-parasitic agents include antibodiesspecific for the parasite.

In some cases, a method of the present disclosure of modulating (e.g.,reducing) an immune response to a microbial parasite modulates an immuneresponse to a microbial parasite such as Plasmodium spp., Toxoplasmagondii, Babesia spp., Trichinella spiralis, Entamoeba histolytica,Giardia lamblia, Enterocytozoon bieneusi, Naegleria, Acanthamoeba,Trypanosoma rhodesiense and Trypanosoma gambiense, Isospora spp.,Cryptosporidium spp, Eimeria spp., Neospora spp., Sarcocystis spp., andSchistosoma spp.

In some cases, a method of the present disclosure of modulating (e.g.,reducing) an immune response to a protozoan parasite modulates an immuneresponse to a protozoan parasite such as Giardia; a plasmodium species(e.g., Plasmodium falciparum); Toxoplasma gondii; a cryptosporidium; aTrichomonas species; a trypanosome (e.g., Trypanosoma cruzi); orLeishmania.

Methods of Modulating an Immune Response to a Pathogenic Fungus

The present disclosure provides methods of modulating an immune responseto a pathogenic fungus, the method comprising administering to anindividual in need thereof an effective amount of an immunomodulatorycomposition of the present disclosure.

In some cases, a method of the present disclosure of modulating animmune response to a pathogenic fungus is effective to reduce the numberof fungal bodies in the individual by at least about 25%, at least about50%, at least about 75%, or at least about 99%, compared to apre-treatment number of fungal bodies in the individual, or to an extentthat the pathogenic fungus cannot be detected in the individual (e.g.,in a biological sample obtained from the individual).

In some cases, a method of the present disclosure of modulating (e.g.,reducing) an immune response to a pathogenic fungus induces or modulatesan immune response to a fungus such as Candida spp. including C.albicans, Aspergillus spp., Cryptococcus spp. including C. neoformans,Blastomyces sp., Pneumocytes spp., or Coccidioides spp.

In some cases, a method of the present disclosure of modulating animmune response to a pathogenic fungus comprises administering animmunomodulatory composition to an individual in need thereof, andfurther comprising administering to the individual an effective amountof a least one additional therapeutic agent. Anti-fungal agents areknown in the art and include, e.g., flucanazole, 5-fluorocytosine, etc.

Suitable anti-fungal agents include, e.g., Polyenes such asAmphotericin-B (including various formulations of Amphotericin-B),Candicidin, Dermostatin, Filipin, Fungichromin, Hachimycin, Hamycin,Lucensomycin, Mepartricin, Natamycin, Nystatin, Pecilocin and Perimycin;and others such as Azaserine, Griseofulvin, Oligomycins, NeomycinUndecylenate, Pyrrolnitrin, Siccanin, Tubercidin and Viridin;Allylamines such as Naftifine and Terbinafine; Imidazoles such asBifonazole, Butoconazole, Chlordantoin, Chlormidazole, Cloconazole,Clotrimazole, Econazole, Enilconazole, Fenticonazole, Isoconazole,Ketoconazole, Miconazole, Omoconazole, Oxiconazole, Nitrate, Sulconazoleand Tioconazole; Triazoles such as Fluconazole, Itraconazole andTerconazole; and other others such as Acrisorcin, Amorolfine,Biphenamine, Bromosalicylchloranilide, Buclosamide, Calcium Propionate,Chlophenesin, Ciclopirox, Cloxyquin, Coparaffinate, Diamthazole,Dihydrochloride, Exalamide, Flucytosine, Halethazole, Hexetidine,Loflucarban, Nifuratel, Potassium Iodide, Propionic Acid, Pyrithione,Salicylanilide, Sodium Propionate, Sulbentine, Tenonitrozole,Tolciclate, Tolindate, Tolnaftate, Tricetin, Ujothion, Undecylenic Acidand Zinc Propionate.

Methods of Treating an Allergic Disease

The present disclosure provides methods of treating an allergic diseasesuch as asthma, allergic rhinitis, conjunctivitis, atopic dermatitis inan individual, the method comprising administering to the individual aneffective amount of an immunomodulatory composition of the presentdisclosure. In some cases, a method of the present disclosure oftreating an allergic disease comprises administering to an individual inneed thereof an effective amount of an immunomodulatory composition ofthe present disclosure, where the immunomodulatory composition comprisesan allergen. Suitable allergens are described above.

In some cases, a subject a method of the present disclosure of treatingan allergic disease is effective to regulate an immune response. In somecases, a subject a method of the present disclosure of treating anallergic disease is effective to decrease one or more of: a) the levelof IgE in an individual; b) the level of allergen-specific IgE in anindividual; c) the number of mast cells in the individual; d) the levelof histamine in the individual; and e) the level of IL-4 in theindividual, compared to a pre-treatment level.

In some cases, a method of the present disclosure of treating anallergic disease comprises administering an immunomodulatory compositionto an individual in need thereof, and further comprising administeringto the individual an effective amount of at least one additionaltherapeutic agent. Suitable additional therapeutic agents include, e.g.,anti-histamines, steroids (e.g., corticosteroids), prostaglandininducers, anti-inflammatory agents, leukotriene antagonists, IL-4muteins, soluble IL-4 receptors, immunosuppressants (such as tolerizingpeptide vaccine), anti-IL-4 antibodies, IL-4 antagonists, anti-IL-5antibodies, soluble IL-13 receptor-Fc fusion proteins, anti-IL-9antibodies, CCR3 antagonists, CCR5 antagonists, VLA-4 inhibitors, anddownregulators of IgE. Suitable steroids include, but are not limitedto, beclomethasone, fluticasone, tramcinolone, budesonide,corticosteroids and budesonide.

Methods of Treating an Autoimmune Disorder

The present disclosure provides methods of treating an autoimmunedisorder in an individual, the method comprising administering to theindividual an effective amount of an immunomodulatory composition of thepresent disclosure. Autoimmune conditions account for many autoimmunedisorders such as rheumatoid arthritis, asthma, type 1 diabetes,multiple sclerosis, systemic lupus erythrymatosus (SLE), Sjorgen'ssyndrome, atherosclerosis, autoimmune hepatitis, autoimmunepancreatitis, celiac disease, autoimmune hemolytic anemia, ankylosingspondylitis, autoimmune disease associated cancers, autoimmune diseaseassociated fibrosis, etc. By modulating innate and adaptive immunemechanisms through the immunomodulatory composition of the presentdisclosure, autoimmune disorders can be treated. In some cases, a methodof the present disclosure of treating an autoimmune disorder comprisesadministering to an individual in need thereof an effective amount of animmunomodulatory composition of the present disclosure, where theimmunomodulatory composition comprises an autoantigen. Suitableautoantigens are described above.

In some cases, a subject a method of the present disclosure of treatingan autoimmune disorder is effective to reduce the number and/or activityof self-reactive T cells in an individual by at least about 25%, atleast about 50%, at least about 75%, or at least about 99% compared to apre-treatment number and/or activity of self-reactive T cells, or to anextent that self-reactive T cells cannot be detected in the individual(e.g., in a biological sample obtained from the individual).

In some cases, a subject a method of the present disclosure of treatingan autoimmune disorder is effective to reduce the level ofautoantibodies in an individual by at least about 25%, at least about50%, at least about 75%, or at least about 99% compared to apre-treatment level of autoantibodies, or to an extent thatautoantibodies cannot be detected in the individual (e.g., in abiological sample obtained from the individual).

In some cases, a subject a method of the present disclosure of treatingan autoimmune disorder is effective to modulate the level of cytokinesin an individual by at least about 25%, at least about 50%, at leastabout 75%, or at least about 99% compared to a pre-treatment level ofcytokines, or to an extent that cytokines cannot be detected in theindividual (e.g., in a biological sample obtained from the individual).

In some cases, a method of the present disclosure of treating anautoimmune disorder comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of a least oneadditional therapeutic agent. Examples of therapeutic agents that can beused to treat autoimmune disorders include, but are not limited to,anti-inflammatory agents; immunosuppressive agents (e.g.,corticosteroids (e.g., prednisone, cortisol, methylprednisolone, etc.)),cyclosporin A); cytotoxic agents (e.g., 6-mercaptopurine, azathioprine,methotrexate, alkylating agents anti-metabolite agents); plantalkaloids; natural products; steroid hormones; hypoxic agents;anti-proliferative agents; anticancer agents; danazol; colchicine;levamisole; biological response modifiers and the like.

Examples of therapeutic agents that can also be used to treat autoimmunedisorders include agents that act to reduce cellular proliferation areknown in the art and widely used. Such agents include alkylating agents,such as nitrogen mustards, nitrosoureas, ethylenimine derivatives, alkylsulfonates, and triazenes, including, but not limited to,mechlorethamine, cyclophosphamide (Cytoxan™), melphalan (L-sarcolysin),carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU),streptozocin, chlorozotocin, uracil mustard, chlormethine, ifosfamide,chlorambucil, pipobroman, triethylenemelamine,triethylenethiophosphoramine, busulfan, dacarbazine, and temozolomide.

Antimetabolite agents include folic acid analogs, pyrimidine analogs,purine analogs, and adenosine deaminase inhibitors, including, but notlimited to, cytarabine (CYTOSAR-U), cytosine arabinoside, fluorouracil(5-FU), floxuridine (FudR), 6-thioguanine, 6-mercaptopurine (6-MP),pentostatin, 5-fluorouracil (5-FU), methotrexate,10-propargyl-5,8-dideazafolate (PDDF, CB3717),5,8-dideazatetrahydrofolic acid (DDATHF), leucovorin, fludarabinephosphate, pentostatine, gemcitabine, cyclocytidine, guanazole, inosineglycodialdehyde, EICAR, ribavirin, tiazofurin, defroxamine andpyrazoloimidazole.

Suitable natural products and their derivatives, (e.g., vinca alkaloids,antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins),include, but are not limited to, Ara-C, paclitaxel (Taxol®), docetaxel(Taxotere®), deoxycoformycin, mitomycin-C, L-asparaginase, azathioprine;brequinar; alkaloids, e.g. vincristine, vinblastine, vinorelbine,vindesine, etc.; podophyllotoxins, e.g. etoposide, teniposide,camptothecin etc.; antibiotics, e.g. anthracycline, daunorubicinhydrochloride (daunomycin, rubidomycin, cerubidine), idarubicin,doxorubicin, epirubicin and morpholino derivatives, etc.; phenoxizonebiscyclopeptides, e.g. dactinomycin; basic glycopeptides, e.g.bleomycin; anthraquinone glycosides, e.g. plicamycin (mithramycin);anthracenediones, e.g. mitoxantrone; azirinopyrrolo indolediones, e.g.mitomycin; macrocyclic immunosuppressants, e.g. cyclosporine, FK-506(tacrolimus, prograf), rapamycin, etc.; antivascular flavonoids; and thelike. Other agents include minerals, nutrients, vitamins, supplements,anti-oxidants, herbs, spices (ginger, oregano, clove etc), naturalhealth products (green tea, fish oil etc) and anti-inflammatorytreatments and modalities.

Other anti-proliferative cytotoxic agents are navelbene, CPT-11,anastrazole, letrazole, capecitabine, reloxafine, cyclophosphamide,folic acid, retinoic acid, ifosamide, and droloxafine. Other suitableanti-proliferative agents include siRNA, interfering RNA (RNAi), andanti-sense RNA.

Microtubule affecting agents that have antiproliferative activity arealso suitable for use and include, but are not limited to,allocolchicine (NSC 406042), Halichondrin B (NSC 609395), colchicine(NSC 757), colchicine derivatives (e.g., NSC 33410), dolstatin 10 (NSC376128), maytansine (NSC 153858), rhizoxin (NSC 332598), paclitaxel(Taxol®), Taxol® derivatives, docetaxel (Taxotere®), thiocolchicine (NSC361792), trityl cysterin, vinblastine sulfate, vincristine sulfate,natural and synthetic epothilones including but not limited to,eopthilone A, epothilone B, discodermolide; estramustine, nocodazole,and the like.

Hormone modulators and steroids (including synthetic analogs) that aresuitable for use include, but are not limited to, adrenocorticosteroids,e.g. prednisone, dexamethasone, etc.; estrogens and pregestins, e.g.hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrolacetate, estradiol, clomiphene, tamoxifen; etc.; and adrenocorticalsuppressants, e.g. aminoglutethimide; 17α-ethinylestradiol;diethylstilbestrol, testosterone, fluoxymesterone, dromostanolonepropionate, testolactone, methylprednisolone, methyl-testosterone,prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone,aminoglutethimide, estramustine, medroxyprogesterone acetate,leuprolide, Flutamide (Drogenil), Toremifene (Fareston), and Zoladex®.Estrogens stimulate proliferation and differentiation; therefore,compounds that bind to the estrogen receptor are used to block thisactivity. Corticosteroids may inhibit T cell proliferation.

Other cytotoxic agents include metal complexes, e.g. cisplatin(cis-DDP), carboplatin, etc.; ureas, e.g. hydroxyurea; and hydrazines,e.g. N-methylhydrazine; epidophyllotoxin; a topoisomerase inhibitor,e.g., irinotecan, etopside phosphate, mitoxantrone; procarbazine;mitoxantrone; leucovorin; tegafur; etc. Other anti-proliferative agentsof interest include immunosuppressants, e.g. mycophenolic acid,thalidomide, desoxyspergualin, azasporine, leflunomide, mizoribine,azaspirane (SKF 105685); Iressa® (ZD 1839,4-β-chloro-4-fluorophenylamino)-7-methoxy-6-β-(4-morpholinyl)propoxy)quinazoline);etc.

Biological response modifiers suitable for use in connection with themethods of the present disclosure include, but are not limited to, (1)inhibitors of tyrosine kinase (RTK) activity; (2) inhibitors ofserine/threonine kinase activity; (3) tumor-associated antigenantagonists, such as antibodies that bind specifically to a tumorantigen; (4) apoptosis receptor agonists; (5) interleukin-2; (6)interferon-α; (7) interferon-γ; (8) colony-stimulating factors; (9)inhibitors of angiogenesis; (10) antagonists of tumor necrosis factor;and (11) BRAF inhibitors.

Methods of Treating Diseases Comprising an Immune Dysregulation

The present disclosure provides methods of modulating, restoring and/orregulating an immune dysfunction in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure. Immunedysfunction conditions account for many diseases such as rheumatoidarthritis (RA) and related diseases, diabetes, psoriasis, systemic lupuserythematosus (SLE) and related diseases, graft-versus-host disease(GVHD), ulcerative colitis, bacterial-induced colitis, Crohn's disease,Alopecia areata, asthma, allergic rhinitis, conjunctivitis, transplantrejection, Hashimoto's thyroiditis, inflammatory bowel diseases (IBD),short bowel syndrome and other gastrointestinal disorders (such asCrohn's disease, ulcerative colitis), cardiovascular diseases, obesity,wound healing, burn recovery, aging, weight gain, fat deposition, etc.Dysregulation of immune responses at the gut mucosal surface can causesystemic immune activation through increased translocation of microbesand microbial products from the intestinal tract. By modulating innateand adaptive immune mechanisms (including immune cells, cytokines,antibodies etc.) through the immunomodulatory composition of the presentdisclosure, immune dysregulation can be prevented and/or treated.

In some cases, a method of the present disclosure of treating an immunedysfunction disorder comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of at least oneadditional therapeutic agent.

In some cases, the method comprising administering to an individual inneed thereof an effective amount of an immunomodulatory composition ofthe present disclosure in a vaccine including an antigen that willmodulate the dysfunctional immune response to a disease related antigen.

In some cases, the method comprising administering to an individual inneed thereof, an effective amount of an immunomodulatory composition ofthe present disclosure in protecting, modulating, restoring orcorrecting disease- or medical condition-related imbalances in thepatients' microbiome. Dysbalance in microbiota accounts for variousdisorders such as dermatological conditions, exuberant inflammatoryresponses, inflammation associated cancers, preterm birth, infertility,female contraception, urogenital infections, sexually transmitteddiseases etc.

Another aspect of the invention includes method of treatment bysubstantially increasing or decreasing a relative abundance ofmiocrobiota of the subject.

In some cases, a method of the present disclosure of treating an immunedysregulation and restoring homeostasis comprises administering animmunomodulatory composition to an individual in need thereof, andfurther comprising administering to the individual an effective amountof one or more members of the microbiome or a probiotic.

Methods of Treating Diseases Comprising an Undesirable InflammatoryActivity

The present disclosure provides methods of modulating and/or regulatingan undesirable inflammatory activity in an individual, the methodcomprising administering to the individual an effective amount of animmunomodulatory composition of the present disclosure. Undesirableinflammatory conditions account for many diseases such as diarrhoealdisease, mucositis due to chemotherapy or radiotherapy, gastroenteritisdue to an infectious agent or an antibiotic agent, pouchitis, obesityrelated inflammation, appendicitis, organ (liver, kidney, lung, heart,islets etc.) transplantation, bacterial infections, viral infections,fungal infections, cancer-associated inflammation, urogenital diseases,bacterial vaginosis, surgical associated trauma, sepsis, anorexia,hyperoxalurea, ulcers, wound healing, renal disease, hepatic diseases,liver fibrosis, alcoholic hepatitis, fibrotic diseases such as lungfibrosis, kidney fibrosis, idiopathic pulmonary fibrosis, acne,undesirable respiratory inflammatory activity, inflammation-associatedcancers, inflammation-associated organ (lung, liver, kidney, heart,gastrointestinal tract, brain etc.) damage and injury, autoinflammatorydiseases (such as TNF receptor associated periodic syndrome, DubinJohnson syndrome, Behcet's disease, familial mediteranean fever etc.)etc. By modulating innate and adaptive immune mechanisms (includingimmune cells, cytokines, chemokines, antibodies etc.) through theimmunomodulatory composition of the present disclosure, inflammatorydisorders can be prevented and/or treated.

In one aspect of the invention, controlling undesirable inflammatoryresponses also include modulation in the levels of hormones,prostaglandins, reactive intermediates and leukotrienes.

In some cases, a method of the present disclosure of treating aninflammatory disorder comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of at least oneadditional therapeutic agent, one or more members of the microbiome or aprobiotic. Examples of therapeutic agents that can be used to treatinflammatory conditions include, but are not limited to,anti-inflammatory agents; immunosuppressive agents; cytotoxic agents andthe like.

Other nonlimiting examples of therapeutic agents that can be used totreat inflammatory conditions include; calcineurin inhibitors (e.g.,pimecrolimus, tacrolimus, etc.), methotrexate, cyclosporine and topicalagents (e.g., tazarotene, anthralin, calciprotriene, corticosteroids,etc.).

In some cases, the method comprising administering to an individual inneed thereof an effective amount of an immunomodulatory composition ofthe present disclosure in a vaccine including an antigen that willmodulate the inflammatory immune response to a disease related antigen.

Other examples of autoimmune diseases, immune dysregulation,inflammation, allergic diseases, dermal diseases, infectious diseases,and organ transplantations for which the composition is useful fortreatment include diseases such as, primary sclerosis polingitis, sprue,autoimmune arthritis, Lyme arthritis, psoriatic arthritis, reactivearthritis, spondyloarthropathy, dermatitis scleroderma, sarcoidosis,disseminated intravascular coagulation, Kawasaki's disease, nephroticsyndrome, chronic fatigue syndrome, fibromyalgia, Wegener'sgranulomatosis, Henoch-Schoenlejn purpurea, microscopic vasculitis ofthe kidneys, chronic active hepatitis, uveitis, septic shock, toxicshock syndrome, sepsis syndrome, cachexia, acquired immunodeficiencysyndrome, acute transverse myelitis, Huntington's chorea, primarybiliary cirrhosis, hemolytic anemia, polyglandular deficiency type Isyndrome and polyglandular deficiency type II syndrome, Schmidt'ssyndrome, adult (acute) respiratory distress syndrome, seronegativearthopathy, arthropathy, Reiter's disease, psoriatic arthropathy,chlamydia, yersinia and salmonella associated arthropathy,spondyloarhopathy, atheromatous disease/arteriosclerosis, allergiccolitis, atopic allergy, food allergies such as peanut allergy, tree nutallergy, egg allergy, milk allergy, soy allergy, wheat allergy, seafoodallergy, shellfish allergy, or sesame seed allergy, autoimmune bullousdisease, pemphigus vulgaris, pemphigus foliaceus, pemphigoid, linear IgAdisease, autoimmune haemolytic anaemia, Coombs positive haemolyticanaemia, acquired pernicious anaemia, juvenile pernicious anaemia,myalgic encephalitis/Royal Free Disease, autoimmune encephalomyelitis,chronic mucocutaneous candidiasis, giant cell arteritis, nonalcoholicfatty liver disease, steatohepatitis, primary sclerosing hepatitis,cryptogenic autoimmune hepatitis, Acquired Immunodeficiency RelatedDiseases, Hepatitis C, common varied immunodeficiency (common variablehypogammaglobulinaemia), dilated cardiomyopathy, fibrotic lung disease,cryptogenic fibrosing alveolitis, postinflammatory interstitial lungdisease, interstitial pneumonitis, connective tissue disease associatedinterstitial lung disease, mixed connective tissue disease associatedlung disease, systemic sclerosis associated interstitial lung disease,Sjogren's disease associated lung disease, ankylosing spondy litisassociated lung disease, vasculitic diffuse lung disease, haemosiderosisassociated lung disease, drug-induced interstitial lung disease,radiation fibrosis, bronchiolitis obliterans, chronic eosinophilicpneumonia, lymphocytic infiltrative lung disease, postinfectiousinterstitial lung disease, gouty arthritis, type-1 autoimmune hepatitis(classical autoimmune or lupoid hepatitis), type-2 autoimmune hepatitis(anti-LKM antibody hepatitis), autoimmune mediated hypoglycemia, type Binsulin resistance with acanthosis nigricans, hypoparathyroidism,osteoarthrosis, primary sclerosing cholangitis, idiopathic leucopenia,autoimmune neutropenia, renal disease NOS, glomerulonephritides,microscopic vasulitis of the kidneys, discoid lupus, erythematosus, maleinfertility idiopathic or NOS, sperm autoimmunity, sympatheticophthalmia, pulmonary hypertension secondary to connective tissuedisease, Goodpasture's syndrome, pulmonary manifestation ofpolyarteritis nodosa, acute rheumatio fever, rheumatoid spondylitis,Still's disease, systemic sclerosis, Takayasu's disease/arteritis,autoimmune thrombocytopenia, idiopathic thrombocytopenia, autoimmunethyroid disease, hyperthyroidism, atrophic autoimmune hypothyroidism,primary myxoedema, phacogenic uveitis, primary vasculitis, vitiligo,anaphylaxis, pet allergies, latex allergies, drug allergies, allergicrhinoconjuctivitis, eosinophilic esophagitis, hypereosinophilicsyndrome, eosinophilic gastroenteritis cutaneous lupus erythematosus,eosinophilic esophagitis, hypereosinophilic syndrome, eosinophilicgastroenteritis, periodontal diseases such as chronic gingivitis andperiodontitis, genitourinary disorders (such as glomerulonephritis,polycystic kidney disease, hydronephrosis, kidney failure, urinary tractobstruction, hyperuricemia etc.), gynaecological disorders (such asvulvodynia, vaginitis, pelvic disorders etc.), reproductive diseases,urological diseases, mitochondria related disorders, pain, migrane,haematological diseases, psychiatric disorders, mouth diseases (such asfoot and mouth disease), musculoskeletol diseases, ocular diseases,renal disorders (such as nephropathic cystinosis), intoxication (such asalcohol intoxication, chronic salicylate intoxication), skin-pruritus,skin-keratosis, skin diseases (such as erythematosquamous, hypertrophicskin disease, popular skin disease, rosacea, pigment disorder, purpura,acne, skin allergy, vitiligo, bullous skin disease, epidermolysis,scleroderma, eczema, cutaneous lymphoma) muscle wasting disease, muscledisorders, bronchus diseases, vascular diseases, uterine fibroids,hormonal imbalance (such as chronic fatigue syndrome), hair loss,osteoporosis, upper respiratory tract infections-associatedinflammation, and Paget's disease.

Methods of Treating Metabolic Disorders

The present disclosure provides methods of treating metabolic disordersin an individual, the method comprising administering to the individualan effective amount of an immunomodulatory composition of the presentdisclosure. Metabolic disorders account for many diseases such asobesity related metabolic dysfunction, diabetes mellitus, insulinresistance, glucose metabolism disorders, hypoinsulinemia,atherosclerosis, hypercholesterolemia, ischemia, metabolic syndrome,oxidative stress, hypertension, endocrine disorders (Addison's disease,Cushing's disease, hyperthyroidism, hypothyroidism, hypopituitarism,polycystic ovary syndrome etc.), abnormal lipid metabolism, obesityrelated disorders (such as bone loss, weight gain etc.), pancreasrelated disorders, mitochondrial disease etc. By modulating innate andadaptive immune mechanisms (including immune cells, cytokines,antibodies etc.) through the immunomodulatory composition of the presentdisclosure, metabolic diseases can be prevented and/or treated.

In some cases, a method of the present disclosure of treating aninflammatory disorder comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of at least oneadditional therapeutic agent, one or more members of microbiome or aprobiotic. For example, therapeutic agents of interest include and arenot limited to those that are anti-inflammatory agents for the treatmentof cardiovascular disease. Such agents include amlodipine, used to lowerblood pressure and prevent chest pain; enalapril, used in the treatmentof hypertension, and some types of chronic heart failure; pravastatin,atorvastatin and rosuvastatin, used for the treatment of dyslipidemiaand the prevention of cardiovascular disease; angiotensin-convertingenzyme (ACE) inhibitors (e.g., benazepril, ramipiril, etc.); angiotensinII receptor blockers (ARBs) (e.g., candesartan, losartan, etc.); betablockers (e.g., acebutolol, bisoprolol, sotalol, etc.); calcium channelblockers (e.g., amlodipine, verapamil, etc.) etc. Other examples oftherapeutic agents of interest include and are not limited to those thatare anti-inflammatory agents for the treatment of diabetes. Such agentsinclude agents that target the IKK—NF-κB pathway; etanercept,infliximab, adalimumab, which target TNF-α; anakinra and canakinumabwhich target IL-1β; tocilizumab which targets IL-6; AMP-activatedprotein kinase activators; sirtuin-1 activators; mammalian target ofrapamycin inhibitors; C—C motif chemokine receptor 2 antagonists, etc.In some cases, therapeutic agents of interest include and are notlimited to those that are anti-inflammtory agents for the treatment ofobesity. Such agents include lorcaserin; Qsymia™ (Vivus); liraglutide,bupropion, naltrexone, lorcaserin, orlistat, phentermine/topiramate etc.

In some cases, therapeutic agents of interest include and are notlimited to those that are used to treat metabolic disorders such asdiabetes include Insulin (e.g., short-, rapid, intermediate andlong-acting insulin), amylinomimetic drugs (e.g., pramlinitide),alpha-glucosidase inhibitors (e.g., acarbose, miglitol, etc.),biguanides (e.g., metformin, etc.), sulfonylureas (e.g., glyburide,glipizide, etc.), meglitinides (e.g., repaglinide), D-phenylalaninederivatives (e.g., nateglinide), thiazolidinediones (e.g.,rosiglitazone, pioglitazone, etc.), DPP-4 inhibitors (e.g. itagliptin,saxagliptin, linagliptin, etc.), glucagon-like receptor-1 (GLP-1)agonists (e.g., exenatide, liraglutide, etc.), sodium glucosetransporter-2 (SGLT-2) inhibitors (e.g., canagliflozin, dapagliflozin,etc.) etc.

Methods of Treating Neurological Disorders

The present disclosure provides methods of modulating and/or regulatingan inflammatory response, the method comprising administering to anindividual in need thereof an effective amount of an immunomodulatorycomposition of the present disclosure. Inflammatory conditions accountfor many neurological disorders such as Alzheimer's, depression,attention deficit hyperactive disorder (ADHD), mood disorders,schizophrenia, multiple sclerosis, Parkinson's disease, autism,Amyotrophic Lateral Sclerosis (ALS), Cerebral malaria disorders,Huntington's disease, anxiety disorders, epilepsy, etc. By modulatinginnate and adaptive immune mechanisms (including immune cells,cytokines, antibodies etc.) through the immunomodulatory composition ofthe present disclosure, neurological disorders can be treated.

In some cases, a method of the present disclosure of treating aneurological disorder comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of at least oneadditional therapeutic agent, one or more members of the microbiome or aprobiotic. Non-limiting examples of disease modifying agents forneurological disorders of interest include, acetylcholinesteraseinhibitors (e.g., donepezil, rivastigmine, etc.), N-methyl, D-aspartatereceptor (NMDAR) antagonists (e.g., memantine, neramexane, etc.),dopaminergic (e.g., carbidopa/Levodopa), dopamine agonists (e.g.pramipexole, ropinirole, apomorphine, etc.), anticholinergics (e.g.,trihexyphenidyl, benztropine mesylate, etc.),catechol-o-methyltransferase (COMT) inhibitors (e.g. entacapone,tolcapone, etc.), anti-convulsants (e.g. diazepam, baclofen, dantrolene,tizanidine, etc.), disease modifying agents; (e.g., teriflunomide,fingolimod, mitoxantrone, dimethyl fumarate, natalizumab, etc.).

In some cases, the method comprising administering to an individual inneed thereof an effective amount of an immunomodulatory composition ofthe present disclosure in a vaccine including an antigen that willmodulate an immune response to a disease related protein such as theamyloid plaques characteristics of Alzheimer or Creutzfeldt-Jacobdisease (CJD).

Methods of Preventing or Treating Immunosuppression and Infections

The present disclosure provides methods of preventing or limiting immunedefect/deficiency/suppression due to viral infections or due toinfections following strokes and other brain injuries comprisingadministering an immunomodulatory composition of the present disclosureto an individual in need thereof. Various forms of viral (e.g., HIV)infections, brain trauma, including stroke, lead to long-term systemicimmune suppression, resulting in higher infection and mortality rates.Further, hepatic invariant NKT cells have been shown to be important toameliorate systemic immunosuppression. The present disclosure representsa strategy to prevent systemic immunosuppression and infections in thesepatients through modulation of NK, NKT and other immune cells.

In some cases, a method of the present disclosure of treating an immunesuppression, stroke or brain trauma disorder comprises administering animmunomodulatory composition to an individual in need thereof, andfurther comprising administering to the individual an effective amountof at least one additional therapeutic agent.

Methods of Enhancing the Efficacy and/or Reducing the Toxicity of aTherapeutic Treatment

The present disclosure also provides methods for enhancing the efficacyand/or reducing the toxicity of a therapeutic treatment, and/orpreventing the drug-resistance and altering metabolism, preferablytreatment with an anti-infective (such as antibacterial, antifungal orantiviral), anticancer agents, therapeutic antibodies, anti-inflammatoryagents, metabolic disorder related drugs, immunostimulatory agents,immunomodulatory compounds, immunoregulatory agents, Si RNAs,therapeutic microbes including probiotics and microbiome (viable,inactivated, heat-killed, mutated, attenuated, genetically engineered)or a surgical treatment by administering an effective amount of animmunomodulatory composition of the present disclosure to an individual,cells or tissues preferably the amount needed to regulate an immuneresponse.

In some cases, a method of the present disclosure of enhancing efficacyand reducing toxicity comprises administering an immunomodulatorycomposition to an individual in need thereof, and further comprisingadministering to the individual an effective amount of at least oneadditional therapeutic agent.

Non-limiting examples of the suitable therapeutic agents and antibodiesare described herein above in methods sections.

Methods of Modulating Dendritic Cells

The present disclosure provides a method of modulating dendritic cells,the method comprising: a) contacting dendritic cells (DCs) obtained froman individual with a composition comprising: i) Caulobacter crescentus;and/or ii) an antigen. The DCs are contacted with the CC and the antigenis in vitro. Contacting DCs with the antigen and the CC modulatesantigen presentation of the antigen on the DCs, thereby generating apopulation of modulated DCs. In some cases, the antigen can be contactedwith DCs using methods such as diffusion, electroporation, activetransport, liposome fusion, phagocytosis, sonication etc. In some cases,the method further comprises administering the antigen-presenting DCs tothe individual from whom the DCs were obtained. In some cases, themethod further comprises administering the antigen-presenting DCscombined with antibodies, chemotherapeutic agents, or cytokines to theindividual from whom the DCs were obtained. Administering modulated DCsto an individual can treat a disease in the individual.

Suitable antigens are described above. In some cases, a compositioncomprising CC and antigen is contacted with DCs; and the CC-antigen-DCmixture is incubated for a period of time of from about 30 minutes toabout 48 hours, thereby generating a population of antigen-presentingDCs. A subject method can modulate the proportion of DCs that areantigen-presenting DCs by at least about 25%, at least about 50%, atleast about 75%, at least about 2-fold, at least about 5-fold, at leastabout 10-fold, at least about 25-fold, at least about 50-fold, at leastabout 100-fold, or more than 100-fold, compared to the proportion of DCsin the starting population that are antigen-presenting DCs.

Methods of Generating Regulatory Immune Cells

The present disclosure provides a method of generating regulatorylymphocytes such as NK, NKT, γδ T cells, ILCs, T cells, and B cells, themethod comprising: a) contacting lymphocytes (NK, NKT, γδ T cells, ILCs,T cells, and/or B cells) obtained from an individual with a compositioncomprising: i) Caulobacter crescentus; and/or ii) an antigen in thepresence or absence of antigen presenting cells. Contacting lymphocytesand the CC generates a population of regulatory lymphocytes. In somecases, the method comprises administering the regulatory lymphocytes tothe individual from whom the cells were obtained, to prevent and/ortreat a disease in a host. In some cases, the method further comprisesadministering the regulatory lymphocytes combined with antibodies,chemotherapeutic agents, or cytokines to the individual from whom thecells were obtained, to prevent and/or treat a disease in a host.

Methods of Treating an Infection with an Intracellular Pathogen

The present disclosure provides methods of preventing and/or treatinginfections with intracellular pathogens (e.g., viruses, mycobacteria,bacteria, parasites etc.) in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition of the present disclosure.

In some cases, a method of the present disclosure of treating anintracellular pathogen comprises administering to an individual in needthereof, and further comprising administering to the individual aneffective amount of at least one additional therapeutic agent.

Methods of Modulating Immune Responses in Animal Models or Cell Culturefor Research, Diagnosis and/or Therapeutic Purposes

The present disclosure provides a method of modulating immune responsesin animal models for research purposes. The present disclosure furtherprovides a method of modulating various TLRs, NLRs, DCs and/or effectorlymphocytes such as NK, NKT, T and B cells, the method comprising: a)contacting effector cells (NK, NKT, T and B cells) obtained from anindividual with a composition comprising: i) Caulobacter crescentus;and/or ii) an antigen in the presence or absence of antigen presentingcells. Contacting effector lymphocytes and the CC modulates theiractivation, thereby generating a population of regulated effectorlymphocytes. In some cases, the method comprises of diagnosing a diseasestate by identifying and expanding specific antigen reactive T cellsand/or B cells. In some cases, the method comprises of identifying andexpanding specific antigen reactive T cells and/or B cells in vitro forresearch purposes. In some cases the method comprises of administeringthe activated effector lymphocytes to the individual from whom the cellswere obtained, to prevent and/or treat a disease in a host. In somecases, the method comprises of activating TLRs or NLRs for researchand/or diagnostic purposes.

Methods of Inducing Proliferation, Differentiation and/or Modulation ofStem Cells

The present disclosure provides a method of inducing proliferation,differentiation and/or modulation of stem cells and restoration ofhomeostasis in an individual, the method comprising administering to theindividual an effective amount of an immunomodulatory composition of thepresent disclosure. The present disclosure provides a method ofmodifying stem cells, the method comprising contacting the stem cellswith a composition comprising Caulobacter crescentus, wherein saidcontacting generates a population of expanded, differentiated and/ormodulated stem cells.

The present disclosure also provides a method of inducing proliferation,differentiation and/or moduation of stem cells, the method comprisingcontacting stem cells obtained from an individual with animmunomodulatory composition of the present disclosure, e.g., animmunomodulatory composition comprising Caulobacter crescentus.Contacting the stem cells with the CC leads to their proliferation anddifferentiation, thereby generating a population of expanded,differentiated and/or modulated cells. The population of expanded,differentiated and/or modulated cells can then be administered to theindividual from whom the stem cells were obtained.

In some embodiments, a method of the present disclosure of inducingproliferation, differentiation and/or modulation of stem cellscomprises: a) obtaining stem cells from an individual; b) contacting thestem cells in vitro with CC, thereby generating a population ofexpanded, differentiated and/or modulated cells; and c) administeringthe population of expanded, differentiated and/or modulated cells to theindividual.

In some embodiments, a method of the present disclosure of inducingproliferation, differentiation and/or modulation of stem cells in anindividual comprises administering to the individual an effective amountof an immunomodulatory composition of the present disclosure. In somecases, an effective amount of an immunomodulatory composition of thepresent disclosure is an amount that is effective, when administered ina single dose or in multiple doses, to induce proliferation,differentiation and/or modulation of hematpoietic stem cells, andrestore homeostasis. For example, in some cases, an effective amount ofan immunomodulatory composition of the present disclosure is an amountthat is effective, when administered in a single dose or in multipledoses, to induce proliferation, differentiation and/or modulation ofhematpoietic stem cells, and restore homeostasis in an individual by atleast 10%, at least 15%, at least 20%, at least 25%, at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 75%, atleast 100% (or 2-fold), at least 2.5-fold, at least 5-fold, at least10-fold, more than 10-fold, at least 15-fold, at least 20-fold, at least25-fold, at least 50-fold, at least 100-fold, or more than 100-fold,compared to the individual in the absence of treatment with theimmunomodulatory composition.

Method of Delivering Therapeutic Molecules

The present disclosure provides a method of delivering therapeuticmolecules such as a protein, peptide, siRNA, carbohydrate,macromolecules etc. where Caulobacter crescentus can act as a carrierand/or delivery vehicle to deliver the therapeutic molecule. As a nongenetic modification (GM), such as electrostatic and hydrophobicinteractions, binding of molecules to the Caulobacter crescentus surfacemay enable the Caulobacter crescentus to act as a carrier and/ordelivery vehicle. Further, due to bioadhesion/mucoadhesion, Caulobactercrescentus may facilitate uptake of the delivered by M cell transport atmucosal surfaces.

Formulations, Dosages, and Routes of Administration

An immunomodulatory composition of the present disclosure can includeone or more pharmaceutically acceptable excipients; and can beformulated in any of a variety of ways, that may depend, e.g., on theroute of administration. Pharmaceutically acceptable excipients areknown to those skilled in the art, and have been amply described in avariety of publications, including, for example, A. Gennaro (1995)“Remington: The Science and Practice of Pharmacy”, 19th edition,Lippincott, Williams, & Wilkins. Suitable excipient vehicles include,for example, water, saline, dextrose, glycerol, ethanol, inert proteins,hydrophillic polymers, amino acids, fatty acids, surfactants, non-ionicsurfactants, carbohydrates, dextrins, polyols, chelating agents, or thelike, and combinations thereof. In addition, if desired, the vehicle maycontain minor amounts of auxiliary substances such as wetting oremulsifying agents or pH buffering agents. Actual methods of preparingsuch dosage forms are known, or will be apparent, to those skilled inthe art. See, e.g., Remington's Pharmaceutical Sciences, Mack PublishingCompany, Easton, Pa., 17th edition, 1985; Remington: The Science andPractice of Pharmacy, A. R. Gennaro, (2000) Lippincott, Williams &Wilkins.

An immunomodulatory composition can be incorporated into a variety offormulations for therapeutic administration. More particularly, animmunomodulatory composition can be formulated into pharmaceuticalcompositions by combination with appropriate, pharmaceuticallyacceptable carriers, salts, preservatives, buffering agents, ordiluents, and may be formulated into preparations in solid, semi-solid,liquid, lyophilized, freeze-dried or gaseous forms, such as tablets,capsules, powders, granules, ointments, solutions, suppositories,injections, skin patches, inhalants and aerosols. In other embodiments,the formulation comprises a colloidal delivery system that includese.g., liposomes, nano-particles, nano-emulsions, nano capsules,microspheres and polymers.

In pharmaceutical dosage forms, an immunomodulatory composition may beadministered alone or in appropriate association, as well as incombination, with other pharmaceutically active compounds. Animmunomodulatory composition, an antigen, adjuvant and/or therapeuticdrug can be administered concurrently, simultaneously, sequentially orat different times, at the same or different sites, and via differentroutes. The following methods and excipients are merely exemplary andare in no way limiting.

For oral preparations, an immunomodulatory composition can be used aloneor in combination with appropriate additives to make tablets, powders,granules or capsules, for example, with conventional additives, such aslactose, mannitol, corn starch or potato starch; with binders, such ascrystalline cellulose, cellulose derivatives, acacia, corn starch orgelatins; with disintegrators, such as corn starch, potato starch orsodium carboxymethylcellulose; with lubricants, such as talc ormagnesium stearate; and if desired, with diluents, buffering agents,moistening agents, preservatives and flavoring agents.

An immunomodulatory composition can be formulated into liquidpreparations for administration by dissolving, suspending or emulsifyingthe composition in an aqueous or nonaqueous solvent, such as vegetableor other similar oils, synthetic aliphatic acid glycerides, esters ofhigher aliphatic acids or propylene glycol; and if desired, withconventional additives such as solubilizers, isotonic agents, suspendingagents, emulsifying agents, stabilizers and preservatives.

An immunomodulatory composition can be utilized in aerosol formulationto be administered via inhalation. The immunomodulatory compositions ofthe present disclosure can be formulated into pressurized acceptablepropellants such as dichlorodifluoromethane, propane, nitrogen and thelike.

Furthermore, an immunomodulatory composition can be made intosuppositories by mixing with a variety of bases such as emulsifyingbases or water-soluble bases. An immunomodulatory composition can beadministered rectally via a suppository. The suppository can includevehicles such as cocoa butter, carbowaxes and polyethylene glycols,which melt at body temperature, yet are solidified at room temperature.

An immunomodulatory composition of the present disclosure can also beadministered in the form of liposomes or liposomal polymeric gels.Liposomes can be given by a variety of routes, oral, nasal, parenteral,trans-dermal, inhalation etc. As is known in the art, liposomes arederived from phospholipids or other lipid substances. Liposomes areformed by mono- or multilamellar hydrated liquid crystals that aredispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto an immunomodulatory composition of the present disclosure, one ormore of a stabilizer, a preservative, an excipients, and the like.Exemplary lipids are the phospholipids and the phosphatidylcholines(lecithins), both natural and synthetic. Liposomes can be in a sizerange of from less than 100 nm to several microns. Methods to formliposomes are known in the art. for example, Prescott, Ed., Methods inCell Biology, Volume XIV, Academic Press, New York, N.Y. (1976), p. 33et seq.

Unit dosage forms for oral or rectal administration such as syrups,elixirs, emulsions, and suspensions may be provided wherein each dosageunit, for example, teaspoonful, tablespoonful, tablet or suppository,contains a predetermined amount of the composition containing one ormore active agents. Similarly, unit dosage forms for injection orintravenous administration may comprise an immunomodulatory compositionas a solution in sterile water, normal saline or anotherpharmaceutically acceptable carrier.

A subject immunomodulatory composition can be formulated for topicaladministration. Topical administration includes administration to theskin or mucosa, including surfaces of the lung eye, nose, and ear.Suitable topical preparations include, e.g., skin patch preparation,transdermal patch preparation, micro arrays, cream, lotion, gelpreparations, powder, ointment, paste, intranasal drops or gels.

Ointments are semi-solid preparations, which are typically based onpetrolatum or other petroleum derivatives. Suitable ointments includeoleaginous bases; emulsifiable bases; emulsion bases; and water-solublebases. Oleaginous ointment bases include, for example, vegetable oils,fats obtained from animals, and semisolid hydrocarbons obtained frompetroleum. Emulsifiable ointment bases, also known as absorbent ointmentbases, contain little or no water and include, for example,hydroxystearin sulfate, anhydrous lanolin and hydrophilic petrolatum.Emulsion ointment bases are either water-in-oil (WIO) emulsions oroil-in-water (OIW) emulsions, and include, for example, cetyl alcohol,glyceryl monostearate, lanolin and stearic acid. Exemplary water-solubleointment bases are prepared from polyethylene glycols of varyingmolecular weight.

Lotions are preparations to be applied to the skin surface withoutfriction, and are typically liquid or semi liquid preparations in whichsolid particles, including the active agent, are present in a water oralcohol base. Lotions are usually suspensions of solids, and preferably,for the present purpose, comprise a liquid oily emulsion of theoil-in-water type. Lotions can be used for treating large body areas,because of the ease of applying a more fluid composition. Lotions maycontain suspending agents to produce better dispersions as well ascompounds useful for localizing and holding the active agent in contactwith the skin, e.g., methyl cellulose, sodium carboxymethyl-cellulose,or the like. An example of a lotion formulation for use in conjunctionwith the present invention contains propylene glycol mixed with ahydrophilic petrolatum such as that which may be obtained under thetrademark Aquaphor® from Beiersdorf, Inc. (Norwalk, Coon.).

Suitable creams can be viscous liquid or semisolid emulsions, eitheroil-in-water or water-in-oil. Cream bases are water-washable, andcontain an oil phase, an emulsifier and an aqueous phase. The oil phase,also sometimes called the “internal” phase, is generally comprised ofpetrolatum and a fatty alcohol such as cetyl or stearyl alcohol; theaqueous phase usually, although not necessarily, exceeds the oil sophase in volume, and generally contains a humectant. The emulsifier in acream formulation, as explained in Remington, supra, is generally anonionic, anionic, cationic or amphoteric surfactant.

Gels formulations can be used. Gels are semisolid, suspension-/typesystems. Single-phase gels contain organic macromolecules distributedsubstantially uniformly throughout the carrier liquid, which can beaqueous, but may also contain an alcohol and, optionally, an oil.

A topical formulation may also be delivered to the skin usingconventional “transdermal”-type patches, wherein the agent(immunomodulatory composition) is contained within a laminated structurethat serves as a delivery device to be affixed to the skin. In such astructure, the immunomodulatory composition is contained in a layer, or“reservoir,” underlying an upper backing layer. The laminated structuremay contain a single reservoir, or it may contain multiple reservoirs.In one embodiment, the reservoir comprises a polymeric matrix of apharmaceutically acceptable contact adhesive material that serves toaffix the system to the skin during drug delivery. Examples of suitableskin contact adhesive materials include, but are not limited to,polyethylenes, polysioxanes, polyisobutylenes, polyacrylates,polyurethanes, and the like. The particular polymeric adhesive selectedwill depend on the particular immunomodulatory composition, vehicle,etc., i.e., the adhesive must be compatible with all components of thedrug-containing composition. In an alternative embodiment, theimmunomodulatory composition-containing reservoir and skin contactadhesive are present as separate and distinct layers, with the adhesiveunderlying the reservoir which, in this case, may be either a polymericmatrix as described above, or it may be a liquid or hydrogel reservoir,or may take some other form.

The term “unit dosage form,” as used herein, refers to physicallydiscrete units suitable as unitary dosages for human and animalsubjects, each unit containing a predetermined quantity of an activeagent (e.g., CC; antigen; etc.) calculated in an amount sufficient toproduce the desired effect in association with a pharmaceuticallyacceptable diluent, carrier or vehicle. The specifications for theactive agents depend on the particular compound employed and the effectto be achieved, and the pharmacodynamics associated with each compoundin the host.

Other modes of administration will also find use. For instance, animmunomodulatory composition can be formulated in suppositories and, insome cases, aerosol and intranasal compositions. For suppositories, thevehicle composition will include traditional binders and carriers suchas, polyalkylene glycols, or triglycerides. Such suppositories may beformed from mixtures containing the active ingredient in the range ofabout 0.5% to about 10% (w/w), or about 1% to about 2%.

Intranasal formulations will usually include vehicles that neither causeirritation to the nasal mucosa nor significantly disturb ciliaryfunction. Diluents such as water, aqueous saline or other knownsubstances can be employed. The nasal formulations may also containpreservatives such as, but not limited to, chlorobutanol andbenzalkonium chloride. A surfactant may be present to enhance absorptionof the subject proteins by the nasal mucosa.

An immunomodulatory composition can be administered as injectables.Typically, injectable compositions are prepared as liquid solutions orsuspensions; solid forms suitable for solution in, or suspension in,liquid vehicles prior to injection may also be prepared. The preparationmay also be emulsified or the active ingredient encapsulated in liposomevehicles.

Suitable excipient vehicles are, for example, water, saline, dextrose,glycerol, ethanol, or the like, and combinations thereof. In addition,if desired, the vehicle may contain minor amounts of auxiliarysubstances such as wetting or emulsifying agents or pH buffering agents.Actual methods of preparing such dosage forms are known, or will beapparent, to those skilled in the art. See, e.g., Remington'sPharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 17thedition, 1985; Remington: The Science and Practice of Pharmacy, A. R.Gennaro, (2000) Lippincott, Williams & Wilkins. The composition orformulation to be administered will, in any event, contain a quantity ofan active agent (e.g., CC; antigen; etc.) adequate to achieve thedesired state in the subject being treated.

The pharmaceutically acceptable excipients, such as vehicles, adjuvants,salts, carriers or diluents, are readily available to the public.Moreover, pharmaceutically acceptable auxiliary substances, such as pHadjusting and buffering agents, tonicity adjusting agents, stabilizers,emulsifying agents, surfactants, preservatives, amino acids, fattyacids, wetting agents and the like, are readily available to the public.

Oral Formulations

In some embodiments, an immunomodulatory composition is formulated fororal delivery to an individual in need of such an immunomodulatorycomposition.

For oral delivery, a subject formulation comprising an immunomodulatorycomposition will in some embodiments include an enteric-soluble coatingmaterial. Suitable enteric-soluble coating material includehydroxypropyl methylcellulose acetate succinate (HPMCAS), hydroxypropylmethyl cellulose phthalate (HPMCP), cellulose acetate phthalate (CAP),polyvinyl phthalic acetate (PVPA), Eudragit™, and shellac.

Suitable oral formulations also include an immunomodulatory composition,formulated with any of the following: microgranules (see, e.g., U.S.Pat. No. 6,458,398); biodegradable macromere (see, e.g., U.S. Pat. No.6,703,037); biodegradable hydrogels (see, e.g., Graham and McNeill(1989) Biomaterials 5:27-36); biodegradable particulate vectors (see,e.g., U.S. Pat. No. 5,736,371); bioabsorbable lactone polymers (see,e.g., U.S. Pat. No. 5,631,015); slow release protein polymers (see,e.g., U.S. Pat. No. 6,699,504; Pelias Technologies, Inc.); apoly(lactide-co-glycolide/polyethylene glycol block copolymer (see,e.g., U.S. Pat. No. 6,630,155; Atrix Laboratories, Inc.); a compositioncomprising a biocompatible polymer and particles of metalcation-stabilized agent dispersed within the polymer (see, e.g., U.S.Pat. No. 6,379,701; Alkermes Controlled Therapeutics, Inc.); andmicrospheres (see, e.g., U.S. Pat. No. 6,303,148; Octoplus, B. V.).

Suitable oral formulations also include an immunomodulatory compositionformulated with any of the following: a carrier such as Emisphere®(Emisphere Technologies, Inc.); TIMERx, a hydrophilic matrix combiningxanthan and locust bean gums which, in the presence of dextrose, form astrong binder gel in water (Penwest); Geminex™ (Penwest); Procise™(GlaxoSmithKline); SAVIT™ (Mistral Pharma Inc.); RingCap™ (Alza Corp.);Smartrix® (Smartrix Technologies, Inc.); SQZgel™ (MacroMed, Inc.);Geomatrix™ (Skye Pharma, Inc.); Oros® Tri-layer (Alza Corporation); andthe like.

Also suitable for use are formulations such as those described in U.S.Pat. No. 6,296,842 (Alkermes Controlled Therapeutics, Inc.); U.S. Pat.No. 6,187,330 (Scion, Inc.); and the like.

Also suitable for use herein are formulations comprising an intestinalabsorption enhancing agent. Suitable intestinal absorption enhancersinclude, but are not limited to, calcium chelators (e.g., citrate,ethylenediamine tetracetic acid); surfactants (e.g., sodium dodecylsulfate, bile salts, palmitoylcarnitine, and sodium salts of fattyacids); toxins (e.g., zonula occludens toxin); and the like.

Suitable oral formulations also include an immunomodulatory composition,formulated as a food supplement (e.g. nutraceuticals, yogurt, frozenyogurt, milk powder, cheese, bars, drinks, prebiotics, symbiotics,paraprobiotics) etc.

Controlled Release Formulations

In some embodiments, an immunomodulatory composition is formulated in acontrolled release formulation.

Controlled release can be taken to mean any one of a number of extendedrelease dosage forms. The following terms may be considered to besubstantially equivalent to controlled release, for the purposes of thepresent invention: continuous release, controlled release, delayedrelease, depot, gradual release, long-term release, programmed release,prolonged release, proportionate release, protracted release,repository, retard, slow release, spaced release, sustained release,time coat, timed release, delayed action, extended action, layered-timeaction, long acting, prolonged action, repeated action, slowing acting,sustained action, sustained-action medications, and extended release.Further discussions of these terms may be found in Lesczek Krowczynski,Extended-Release Dosage Forms, 1987 (CRC Press, Inc.).

The various controlled release technologies cover a very broad spectrumof drug dosage forms. Controlled release technologies include, but arenot limited to physical systems and chemical systems.

Physical systems include, but are not limited to, reservoir systems withrate-controlling membranes, such as microencapsulation,macroencapsulation, and membrane systems; reservoir systems withoutrate-controlling membranes, such as hollow fibers, ultra microporouscellulose triacetate, and porous polymeric substrates and foams;monolithic systems, including those systems physically dissolved innon-porous, polymeric, or elastomeric matrices (e.g., nonerodible,erodible, environmental agent ingression, and degradable), and materialsphysically dispersed in non-porous, polymeric, or elastomeric matrices(e.g., nonerodible, erodible, environmental agent ingression, anddegradable); laminated structures, including reservoir layers chemicallysimilar or dissimilar to outer control layers; and other physicalmethods, such as osmotic pumps, or adsorption onto ion-exchange resins.

Chemical systems include, but are not limited to, chemical erosion ofpolymer matrices (e.g., heterogeneous, or homogeneous erosion), orbiological erosion of a polymer matrix (e.g., heterogeneous, orhomogeneous). Additional discussion of categories of systems forcontrolled release may be found in Agis F. Kydonieus, Controlled ReleaseTechnologies: Methods, Theory and Applications, 1980 (CRC Press, Inc.).

There are a number of controlled release drug formulations that aredeveloped for oral administration. These include, but are not limitedto, osmotic pressure-controlled gastrointestinal delivery systems;hydrodynamic pressure-controlled gastrointestinal delivery systems;membrane permeation-controlled gastrointestinal delivery systems, whichinclude microporous membrane permeation-controlled gastrointestinaldelivery devices; gastric fluid-resistant intestine targetedcontrolled-release gastrointestinal delivery devices; geldiffusion-controlled gastrointestinal delivery systems; andion-exchange-controlled gastrointestinal delivery systems, which includecationic and anionic drugs. Additional information regarding controlledrelease drug delivery systems may be found in Yie W. Chien, Novel DrugDelivery Systems, 1992 (Marcel Dekker, Inc.). Some of these formulationswill now be discussed in more detail.

Enteric coatings are applied to tablets to prevent the release of activeagents in the stomach either to reduce the risk of unpleasant sideeffects or to maintain the stability of the drug which might otherwisebe subject to degradation of expose to the gastric environment. Mostpolymers that are used for this purpose are polyacids that function byvirtue or the fact that their solubility in aqueous medium ispH-dependent, and they require conditions with a pH higher than normallyencountered in the stomach.

One exemplary type of oral controlled release structure is entericcoating of a solid or liquid dosage form. The enteric coatings aredesigned to disintegrate in intestinal fluid for ready absorption. Delayof absorption of the active agent that is incorporated into aformulation with an enteric coating is dependent on the rate of transferthrough the gastrointestinal tract, and so the rate of gastric emptyingis an important factor. Some investigators have reported that amultiple-unit type dosage form, such as granules, may be superior to asingle-unit type.

Suitable enteric coating agents include, but are not limited to,hydroxypropylmethylcellulose phthalate, methacryclic acid-methacrylicacid ester copolymer, polyvinyl acetate-phthalate and cellulose acetatephthalate.

Another type of useful oral controlled release structure is a soliddispersion. A solid dispersion may be defined as a dispersion of one ormore active ingredients in an inert carrier or matrix in the solid stateprepared by the melting (fusion), solvent, or melting-solvent method.

Examples of carriers useful in solid dispersions include, but are notlimited to, water-soluble polymers such as polyethylene glycol,polyvinylpyraolidone, and hydroxypropylmethylcellulose. Alternativecarriers include phosphatidylcholine. Phosphatidylcholine is anamphoteric but water-insoluble lipid, which may improve the solubilityof otherwise insoluble active agents in an amorphous state inphosphatidylcholine solid dispersions.

Other carriers include polyoxyethylene hydrogenated castor oil. Animmunomodulatory composition can be included in a solid dispersionsystem with an enteric polymer such as hydroxypropylmethylcellulosephthalate and carboxymethylethylcellulose, and a non-enteric polymer,hydroxypropylmethylcellulose. Another solid dispersion dosage formincludes incorporation of the drug of interest (e.g., an active agent)with ethyl cellulose and stearic acid in different ratios.

There are various methods commonly known for preparing soliddispersions. These include, but are not limited to, the melting method,the solvent method and the melting-solvent method.

Injectable microspheres are another controlled release dosage form.Injectable micro spheres may be prepared by non-aqueous phase separationtechniques, and spray-drying techniques. Microspheres may be preparedusing polylactic acid or copoly(lactic/glycolic acid).

Other controlled release technologies that may be used include, but arenot limited to, SODAS (Spheroidal Oral Drug Absorption System), INDAS(Insoluble Drug Absorption System), IPDAS (Intestinal Protective DrugAbsorption System), MODAS (Multiporous Oral Drug Absorption System),EFVAS (Effervescent Drug Absorption System), PRODAS (Programmable OralDrug Absorption System), and DUREDAS (Dual Release Drug AbsorptionSystem) available from Elan Pharmaceutical Technologies. SODAS are multiparticulate dosage forms utilizing controlled release beads. INDAS are afamily of drug delivery technologies designed to increase the solubilityof poorly soluble drugs. IPDAS are multi particulate tablet formationutilizing a combination of high density controlled release beads and animmediate release granulate. MODAS are controlled release single unitdosage forms. Each tablet consists of an inner core surrounded by asemipermeable multiparous membrane that controls the rate of drugrelease. EFVAS is an effervescent drug absorption system. PRODAS is afamily of multi particulate formulations utilizing combinations ofimmediate release and controlled release mini-tablets. DUREDAS is abilayer tablet formulation providing dual release rates within the onedosage form. Although these dosage forms are known to one of skill,certain of these dosage forms will now be discussed in more detail.

An immunomodulatory composition of the present disclosure can beincorporated into any one of the aforementioned controlled releaseddosage forms, or other conventional dosage forms. The amount of activeagent contained in each dose can be adjusted, to meet the needs of theindividual patient, and the indication. One of skill in the art andreading this disclosure will readily recognize how to adjust the levelof an active agent and the release rates in a controlled releaseformulation, in order to optimize delivery of an active agent and itsbioavailability.

Inhalational Formulations

An immunomodulatory composition of the present disclosure will in someembodiments be administered to a patient by means of a pharmaceuticaldelivery system for the inhalation route. The immunomodulatorycomposition may be formulated in a form suitable for administration byinhalation. The inhalational route of administration provides theadvantage that the inhaled drug can bypass the blood-brain barrier. Thepharmaceutical delivery system is one that is suitable for respiratorytherapy by delivery of an active agent to mucosal linings of thebronchi. A system that depends on the power of a compressed gas to expelthe immunomodulatory composition from a container can also be used. Anaerosol or pressurized package can be employed for this purpose.

As used herein, the term “aerosol” is used in its conventional sense asreferring to very fine liquid or solid particles carries by a propellantgas under pressure to a site of therapeutic application. When apharmaceutical aerosol is employed, the aerosol contains thetherapeutically active compound (e.g., active agent), which can bedissolved, suspended, or emulsified in a mixture of a fluid carrier anda propellant. The aerosol can be in the form of a solution, suspension,emulsion, powder, or semi-solid preparation. Aerosols can be used foradministration as fine, solid particles or as liquid mists via therespiratory tract of a patient. Various types of propellants known toone of skill in the art can be utilized. Suitable propellants include,but are not limited to, hydrocarbons or other suitable gas. In the caseof the pressurized aerosol, the dosage unit may be determined byproviding a value to deliver a metered amount.

An immunomodulatory composition can also be formulated for delivery witha nebulizer, which is an instrument that generates very fine liquidparticles of substantially uniform size in a gas. For example, a liquidcontaining the immunomodulatory composition is dispersed as droplets.The small droplets can be carried by a current of air through an outlettube of the nebulizer. The resulting mist penetrates into therespiratory tract of the patient.

There are several different types of inhalation methodologies which canbe employed in connection with an immunomodulatory composition of thepresent disclosure. An immunomodulatory composition can be formulatedwith low boiling point propellants. Such formulations are generallyadministered by conventional meter dose inhalers (MDI's). Alternatively,immunomodulatory composition can be formulated in aqueous or ethanolicsolutions and delivered by conventional nebulizers. In some embodiments,such solution formulations are aerosolized using devices and systemssuch as disclosed within U.S. Pat. Nos. 5,497,763; 5,544,646; 5,718,222;and 5,660,166. An immunomodulatory composition can be formulated intodry powder formulations. Such formulations can be administered by simplyinhaling the dry powder formulation after creating an aerosol mist ofthe powder. Technology for carrying such out is described within U.S.Pat. No. 5,775,320 issued Jul. 7, 1998 and U.S. Pat. No. 5,740,794issued Apr. 21, 1998.

An immunomodulatory composition of the present disclosure will in someembodiments be formulated for vaginal delivery. A subjectimmunomodulatory composition for intravaginal administration can beformulated as an intravaginal bioadhesive tablet, intravaginalbioadhesive microparticle, intravaginal cream, intravaginal lotion,intravaginal foam, intravaginal ointment, intravaginal paste,intravaginal solution, or intravaginal gel.

A subject immunomodulatory composition will in some embodiments beformulated for rectal delivery. A subject formulation for intrarectaladministration comprises a subject immunomodulatory compositionformulated as an intrarectal bioadhesive tablet, intrarectal bioadhesivemicroparticle, intrarectal cream, intrarectal lotion, intrarectal foam,intrarectal ointment, intrarectal paste, intrarectal solution, orintrarectal gel. An immunomodulatory composition of the presentdisclosure can be formulated with agents that improve adhesion tomucosal membranes such as mucoadhesives, bioadhesives, particles,microspheres or liposomes.

A subject immunomodulatory composition can include one or more of anexcipient (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose,cellulose, talc, calcium phosphate or calcium carbonate), a binder(e.g., cellulose, methylcellulose, hydroxymethylcellulose,polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic,poly(ethylene glycol), sucrose or starch), a disintegrator (e.g.,starch, carboxymethylcellulose, hydroxypropyl starch, low substitutedhydroxypropylcellulose, sodium bicarbonate, calcium phosphate or calciumcitrate), a lubricant (e.g., magnesium stearate, light anhydrous silicicacid, talc or sodium lauryl sulfate), a flavoring agent (e.g., citricacid, menthol, glycine or orange powder), a preservative (e.g., sodiumbenzoate, sodium bisulfite, methylparaben or propylparaben), astabilizer (e.g., citric acid, sodium citrate or acetic acid), asuspending agent (e.g., methylcellulose, polyvinylpyrrolidone oraluminum stearate), a dispersing agent (e.g.,hydroxypropylmethylcellulose), a diluent (e.g., water), and base wax(e.g., cocoa butter, white petrolatum or polyethylene glycol).

Tablets comprising an immunomodulatory composition may be coated with asuitable film-forming agent, e.g., hydroxypropylmethyl cellulose,hydroxypropyl cellulose or ethyl cellulose, to which a suitableexcipient may optionally be added, e.g., a softener such as glycerol,propylene glycol, diethylphthalate, or glycerol triacetate; a fillersuch as sucrose, sorbitol, xylitol, glucose, or lactose; a colorant suchas titanium hydroxide; and the like.

Dosages

The dosage of an immunomodulatory composition of the present disclosurecan vary, depending on factors such as the clinical goals to beachieved, the age of the individual being treated, the physical statusof the individual being treated, etc.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10³ CC per unit dosage form to about 10²⁰CC per unit dosage form. For example, an immunomodulatory composition ofthe present disclosure can comprise CC in an amount of from about 10³ CCper unit dosage form to about 10⁴ CC per unit dosage form, from about10⁴ CC per unit dosage form to about 10⁵ CC per unit dosage form, fromabout 10⁵ CC per unit dosage form to about 10⁶ CC per unit dosage form,from about 10⁶ CC per unit dosage form to about 10⁷ CC per ml, fromabout 10⁸ CC per unit dosage form to about 10⁹ CC per unit dosage form,from about 10⁹ CC per ml to about 10¹⁰ CC per unit dosage form, fromabout 10¹⁵ CC per unit dosage form to about 10²⁰ CC per unit dosageform, or more than 10²⁰ CC per unit dosage form.

For example, an immunomodulatory composition of the present disclosurecan comprise CC in an amount of from about 10³ CC per ml to about 10²⁰CC per ml. For example, an immunomodulatory composition of the presentdisclosure can comprise CC in an amount of from about 10³ CC per ml toabout 10⁴ CC per ml, from about 10⁴ CC per ml to about 10⁵ CC per ml,from about 10⁵ CC per ml to about 10⁶ CC per ml, from about 10⁶ CC perml to about 10⁷ CC per ml, from about 10⁸ CC per ml to about 10⁹ CC perml, from about 10⁹ CC per ml to about 10¹⁰ CC per ml, from about 10¹⁵ CCper ml to about 10²⁰ CC per ml, or more than 10²⁰ CC per ml.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10² to about 10²⁰ colony forming units(cfu) per unit dosage form; for example, an immunomodulatory compositionof the present disclosure can comprise CC in an amount of from about 10²to about 10³ from about 10³ to about 10⁵, from about 10⁵ to about 10⁷,from about 10⁷ to about 10⁹, from about 10⁹ to about 10¹¹, from about10¹¹ to about 10¹³, from about 10¹³ to about 10¹⁵, from about 10¹⁵ toabout 10¹⁸, or from about 10¹⁸ to about 10²⁰, cfu per unit dosage form.A unit dosage form can be an amount that is administered in a singledose; for example, a unit dosage form can be 0.5 ml, 1.0 ml, or othervolume suitable for administration in a single dose.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10⁸ CC per mg to about 10¹² CC per mg. Forexample, an immunomodulatory composition of the present disclosure cancomprise CC in an amount of from about 10⁸ CC per mg to about 10⁴ CC permg, from about 10⁴ CC per mg to about 10⁵ CC per mg, from about 10⁵ CCper mg to about 10⁶ CC per mg, from about 10⁶ CC per mg to about 10⁷ CCper mg, from about 10⁸ CC per mg to about 10⁹ CC per mg, from about 10⁹CC per mg to about 10¹⁰ CC per mg, from about 10¹⁰ CC per mg to about10¹¹ CC per mg, or from about 10¹¹ CC per mg to about 10¹² CC per mg.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10³ CC per gram to about 10¹⁵ CC per gram.For example, an immunomodulatory composition of the present disclosurecan comprise CC in an amount of from about 10³ CC per gram to about 10⁴CC per gram, from about 10⁴ CC per gram to about 10⁵ CC per gram, fromabout 10⁵ CC per gram to about 10⁶ CC per gram, from about 10⁶ CC pergram to about 10⁷ CC per gram, from about 10⁸ CC per gram to about 10⁹CC per gram, from about 10⁹ CC per gram to about 10¹⁰ CC per gram, fromabout 10¹⁰ CC per gram to about 10¹¹ CC per gram, from about 10¹¹ CC pergram to about 10¹² CC per gram, from about 10¹² CC per gram to about10¹³ CC per gram, from about 10¹³ CC per gram to about 10¹⁴ CC per gram,or from about 10¹⁴ CC per gram to about 10¹⁵ CC per gram.

An immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10² to about 10²⁰ cfu per ml; for example,an immunomodulatory composition of the present disclosure can compriseCC in an amount of from about 10² to about 10⁸ from about 10⁸ to about10⁵, from about 10⁵ to about 10⁷, from about 10⁷ to about 10⁹, fromabout 10⁹ to about 10″, from about 10″ to about 10¹³, from about 10¹³ toabout 10¹⁵, from about 10¹⁵ to about 10¹⁸, or from about 10¹⁸ to about10²⁰, cfu per ml.

In some embodiments, multiple doses of an immunomodulatory compositionof the present disclosure are administered. The frequency ofadministration of an immunomodulatory composition of the presentdisclosure can vary depending on any of a variety of factors, e.g.,severity of the symptoms, etc. For example, in some embodiments, animmunomodulatory composition of the present disclosure is administeredonce per month, twice per month, three times per month, every other week(qow), once per week (qw), twice per week (biw), three times per week(tiw), four times per week, five times per week, six times per week,every other day (qod), daily (qd), twice a day (qid), or three times aday (tid).

The duration of administration of an immunomodulatory composition of thepresent disclosure, e.g., the period of time over which animmunomodulatory composition of the present disclosure is administered,can vary, depending on any of a variety of factors, e.g., patientresponse, etc. For example, an immunomodulatory composition of thepresent disclosure can be administered over a period of time rangingfrom about one hour to one day, from about one day to about one week,from about two weeks to about four weeks, from about one month to abouttwo months, from about two months to about four months, from about fourmonths to about six months, from about six months to about eight months,from about eight months to about 1 year, from about 1 year to about 2years, or from about 2 years to about 4 years, or more.

Where an immunomodulatory composition comprises an antigen, the dosageof antigen is selected as an amount which is effective and modulates animmune response without significant adverse side effects. Such amountcan vary, depending, e.g., upon which specific antigen is employed, theroute of administration, etc. Where an immunomodulatory compositioncomprises an antigen, the dosage of antigen can range from 1 ng per unitdosage form to about 100 mg per unit dosage form, e.g., from about 1 ngto about 25 ng, from about 25 ng to about 50 ng, from about 50 ng toabout 100 ng, from about 100 ng to about 250 ng, from about 250 ng toabout 500 ng, from about 500 ng to about 750 ng, from about 750 ng toabout 1 μg, from about 1 μg to about 25 ng, from about 25 μg to about 50ng, from about 50 μg to about 100 ng, from about 100 μg to about 250 ng,from about 250 μg to about 500 μg, from about 500 μg to about 750 μg,from about 750 μg to about 1 mg, from about 1 mg to about 25 mg, fromabout 25 mg to about 50 mg, or from about 50 mg to about 100 mg, perunit dosage form.

Routes of Administration

An immunomodulatory composition of the present disclosure isadministered to an individual using any available method and routesuitable for drug delivery, including in vivo and ex vivo methods, aswell as systemic and localized routes of administration.

Conventional and pharmaceutically acceptable routes of administrationinclude intranasal, intramuscular, intratracheal, subcutaneous,intradermal, intranodal, percutaneous, transdermal, intratumoral,topical application, intravenous, intravesicular, rectal, nasal, oraland other enteral and parenteral routes of administration. Routes ofadministration may be combined, if desired, or adjusted depending uponthe agent and/or the desired effect. The composition can be administeredin a single dose or in multiple doses.

An immunomodulatory composition of the present disclosure can beadministered to a host using any available conventional methods androutes suitable for delivery of conventional drugs, including systemicor localized routes. In general, routes of administration contemplatedinclude, but are not necessarily limited to, enteral, parenteral, orinhalational routes.

Parenteral routes of administration other than inhalation administrationinclude, but are not necessarily limited to, topical, transdermal,subcutaneous, intramuscular, intradermal, intralymphatic, intraorbital,intracapsular, intraspinal, intrasternal, intracranial, intravesicular,and intravenous routes, i.e., any route of administration other thanthrough the alimentary canal. Parenteral administration can be carriedto effect systemic or local delivery of the immunomodulatorycomposition. Where systemic delivery is desired, administrationtypically involves invasive or systemically absorbed topical or mucosaladministration of pharmaceutical preparations.

An immunomodulatory composition of the present disclosure can also bedelivered to the subject by enteral administration. Enteral routes ofadministration include, but are not necessarily limited to, oral andrectal (e.g., using a suppository) delivery.

An immunomodulatory composition of the present disclosure can also bedelivered to the subject via a mucosal route of delivery. Mucosal routesof delivery include nasal, buccal, sublingual, vaginal, ocular, andrectal routes of administration.

In certain embodiments, an immunomodulatory composition of the presentdisclosure is administered to a subject via a combination of differentroutes in the order indicated below:

-   -   i. systemic, mucosal;    -   ii. systemic, systemic, mucosal, mucosal;    -   iii. systemic, mucosal, systemic;    -   iv. mucosal, mucosal, systemic, systemic;    -   v. mucosal, systemic, systemic;    -   vi. mucosal, systemic, mucosal, for example.

When an immunomodulatory composition of the present disclosure isadministered systemically or mucosally more than once, the two or moresystemic or mucosal administrations may be by the same systemic (forexample, two intramuscular injections) or mucosal route (two IN/SLadministrations) or different (for example, one intramuscular injectionand one intravenous injection; one IN administration and one SLadministration).

An immunomodulatory composition of the present disclosure isadministered to an individual using any available method, delivery ordevice such as vaccine patches, needles, microneedles (hollow or solid),drop, syrup, tablets, capsules, pipette, dose-spray pumps, nasaldropper, inhalation devices, liquid or dry powder, suspensions orsolutions, spray devices, Accuspray, thermoresponsive gels, jetinjectors, Nasovak, Bespak, ointment, lotions, suppositories, gels etc.

Suitable routes of administration are known in the art; any known routeof administration can be employed in connection with administering animmunomodulatory composition of the present disclosure. See, e.g.,Nursing Drug Guide: Nursing Drug Handbook (2015) 36^(th) ed, Lippincott.

Individuals Suitable for Treatment

Individuals suitable for treatment using a method of the presentdisclosure include humans; non-human mammals; fish; and birds. In any ofthe above embodiments discussed below, the individual being treatedusing a subject method can be a non-human mammal such as livestock(e.g., pigs, sheep, goats, cattles, equine, caprine, ovine, bovine,etc.); a mammalian pet (e.g., cats; dogs; horses; etc.); a bird such aschicken, hens, turkeys, geese, quail, ducks etc.; or other animals suchas fish.

In any of the above embodiments discussed below, the individual beingtreated using a subject method is a human of from about one month toabout 6 months, from about 6 months to about 1 year, or from about 1year to about 5 years of age. In any of the above embodiments discussedbelow, the individual being treated using a subject method is a human offrom about 5 years to about 12 years, from about 13 years to about 18years, or from about 18 years to about 25 years of age. In any of theabove embodiments discussed below, the individual being treated using asubject method is a human of from about 25 years to about 50 years, fromabout 50 years to about 75 years of age, or older than 75 years of age.In any of the above embodiments discussed below, the individual beingtreated using a subject method is a human who is immunocompromised.

In some embodiments, the individual has a viral disease, or is at riskof contracting a viral disease. In some cases, the disease is a viraldisease selected from the group consisting of, but not limited to, viraldisease caused by Zika virus, hepatitis B, hepatitis C, rotavirus, humanimmunodeficiency virus, human T-cell lymphotropic virus, DNA virusessuch as parvoviruses, adeno viruses, papovaviruses (e.g., papillomavirus, polyoma viruses, and SV40), herpes viruses (e.g., herpes simplextype I (HSV-I), herpes simplex type II (HSV-II), and Epstein-Barrvirus), poxviruses (e.g., variola (smallpox) and vaccinia virus); andRNA viruses, such as retroviruses [e.g. human immunodeficiency virustype I (HIV-I), human immunodeficiency virus type II (HIV-II), humanT-cell lymphotropic virus type I (HTLV-I), human T-cell lymphotropicvirus type II (HTLV-II)], orthomyxoviruses (e.g., influenza viruses),paramyxoviruses (e.g., measles virus, mumps virus, respiratory syncytialvirus), rhabdoviruses (e.g., rabies virus), Sendai virus, picornaviruses(e.g., poliomyelitis virus, coxsackieviruses, rhinoviruses), reoviruses(e.g., rotavirus, colorado tick fever virus), togaviruses (e.g., rubellavirus (German measles), Japanese encephalitis virus and Semliki forestvirus), arboviruses, calciviruses (e.g., hepatitis E virus),flaviviruses (e.g., yellow fever virus, dengue virus), coronaviruses,filoviruses (e.g., Ebola and Marburg viruses) and Bunyaviruses (e.g.,Hanta virus, California encephalitis virus).

In some embodiments, the individual has a bacterial infection, or is arisk of contracting a bacterial infection. In some embodiments, theindividual has a mycobacterial infection, or is at risk of contracting amycobacterial infection. In some embodiments, the individual is infectedwith, or is at risk of becoming infected with, a pathogenic bacterium.Pathogenic bacteria include, e.g., Gram positive bacteria, Gram negativebacteria, mycobacteria, etc. Non-limiting examples of pathogenicbacteria include Mycobacteria (e.g., M. tuberculosis, M. avium complex),nontuberculosis Mycobacteria, Streptococcus, Staphylococcus,Pseudomonas, Salmonella, Neisseria, and Listeria. In some cases, thebacteria is Neisseria gonorrhea, M. tuberculosis, M. leprae, Listeriamonocytogenes, Streptococcus pneumoniae, S. pyogenes, S. agalactiae, S.viridans, S. faecalis, or S. Bovis. Other examples of pathogenicbacteria contemplated include, but are not limited to, Gram positivebacteria (e.g., Listeria, Bacillus such as Bacillus anthracia,Erysipelothrix species), Gram negative bacteria (e.g., Bartonella,Brucella, Campylobacter, Enterobacter, Escherichia, Francisella,Hemophilus, Klebsiella, Morganella, Proteus, Providencia, Pseudomonas,Salmonella, Serratia, Shigella, Vibrio, and Yersinia species),spirochete bacteria (e.g., Borrelia species including Borreliaburgdorferi that causes Lyme disease), anaerobic bacteria (e.g.,Actinomyces and Clostridium species), Gram positive and negative coccalbacteria, Enterococcus species, Streptococcus species, Pneumococcusspecies, Staphylococcus species, Neisseria species.

In some cases, the individual has, or is at risk of contracting, aparasitic disease. Parasitic diseases that can be treated or preventedby the methods of the present disclosure include, but are not limitedto, amebiasis, malaria, leishmania, coccidia, giardiasis,cryptosporidiosis, toxoplasmosis, trypanosomiasis, schistosomiasis, andfilariasis.

In some cases, the individual has, or is at risk of contracting, afungal disease. Fungal diseases that can be treated or prevented by themethods of the present disclosure include, but are not limited toCandida spp. including C. albicans, Aspergillus spp., Cryptococcus spp.including C. neoformans, Blastomyces sp., Pneumocytes spp., yeast, mold,or Coccidioides spp.

In some cases, the individual has, or is at risk of contracting, a worminfection, a fluke infection, etc. Also encompassed are infections byvarious worms, such as but not limited to ascariasis, ancylostomiasis,trichuriasis, strongyloidiasis, toxoccariasis, trichinosis,onchocerciasis filaria, and dirofilariasis. Also encompassed areinfections by various flukes, such as but not limited toschistosomiasis, paragonimiasis, and clonorchiasis.

In some embodiments, the individual has an autoimmune disorder,inflammatory disorder or an immune dysfunction, or is at risk ofdeveloping an autoimmune disorder, inflammatory disorder or an immunedysfunction. In some cases, the disease is selected from the groupconsisting of, but not limited to, allergy, rheumatoid arthritis,asthma, diabetes, systemic lupus erythematosus (SLE), Grave's disease,atherosclerosis, multiple sclerosis, schizophrenia, Alzheimer's,depression, hypopituitarism, neurodegenerative disorders, cardiovasculardiseases, obesity, organ transplantation, sepsis, hepatic diseases,psoriasis, metabolic diseases, etc.

In some cases, the disease is selected from the group consisting ofautoimmune and autoimmune-related diseases, including, but not limitedto, acute disseminated encephalomyelitis, acute necrotizing hemorrhagicleukoencephalitis, Addison's disease, agammaglobulinemia, alopeciaareata, amyloidosis, ankylosing spondylitis, anti-GBM/anti-TBMnephritis, antiphospholipid syndrome, angioedema, aplastic anemia,dysautonomia, hepatitis, hyperlipidemia, immunodeficiency, inner eardisease, myocarditis, oophoritis, pancreatitis, retinopathy,thrombocytopenic purpura, thyroid disease, urticarial, axonal andneuronal neuropathies, Balo disease, Behcet's disease, bullouspemphigoid, cardiomyopathy, Castleman disease, celiac disease, Chagasdisease, chronic fatigue syndrome, chronic inflammatory demyelinatingpolyneuropathy, chronic recurrent multifocal osteomyelitis,Churg-Strauss syndrome, cicatricial pemphigoid/benign mucosalpemphigoid, Crohn's disease, Cogans syndrome, cold allutinin disease,congenital heart block, Coxsacke myocarditis, CREST disease, essentialmixed cryoglobulinemia, demyelinating neuropathies, dermatitisherpetiformis, dermatomyositis, Devic's disease, discoid lupus,Dressler' syndrome, endometriosis, eosinophilic esophagitis,eosinophilic fasciitis, erythema nodosum, experimental allergicencephalomyelitis, Evans syndrome, fibromyalgia, fibrosing alveolitis,giant cell arteritis, giant cell myocarditis, glomerulonephritis,Goodpasture's syndrome, granulomatosis with polyangiitis, Graves'disease, Guillain-Barre syndrome, Hashimoto's encephalitis, Hashimoto'sthyroiditis, hemolytic anemia, Henoch-Schonlein purpura, herpesgestationis, hypogammaglobulinemia, idiopathic thrombocytopenic purpura,IgA nephropathy, IgG4-related sclerosing disease, immunoregulatorylipoproteins, inclusion body myositis, interstitial cystitis, juvenialarthritis, juvenial diabetes (Type 1 diabetes), juvenile myositis,Kawasaki syndrome, Lamber-Eaton syndrome, leukocytoclastic vasculitis,lichen planus, lichen sclerosus, ligneous conjunctivitis, linear IgAdisease, lupus, lyme disease, Meniere's disease, microscopicpolyangiitis, mixed connective tissue disease, Mooren's ulcer,Mucha-Habermann disease, multiple sclerosis, myasthenia gravis,myostitis, narcolepsy, neuromyelitis optica, neutropenia, ocularcicatricial pemphigoid, optic neuritis, palindromic rheumatism, PANDAS,paraneoplastic cerebellar degeneration, paroxysmal nocturnalhemoglobinuria, Parry Romber syndrome, Parsonnage-Turner syndrome, parsplanitis, pemphigus, peripheral neuropathy, perivenousencephalomyelitis, pernicious anemia, POEMS syndrome, polyarteritisnodosa, Type I, II and III autoimmune polyglandular syndromes,polymyalgia rheumatic, polymyositis, postmyocardial infarction syndrome,postpericardiotomy syndrome, progesterone dermatitis, primary biliarycirrhosis, primary sclerosing cholangitis, psoriasis, psoriaticarthritis, idiopathic pulmonary fibrosis, pyoderma gengrenosum, pure redcell aplasia, Raynauds phenomenon, reactive arthritis, reflexsympathetic dystrophy, Reiter's syndrome, relapsing polychondritis,restless legs syndrome, retroperitoneal fibrosis, rheumatic fever,rheumatoid arthritis, sarcoidosis, Schmidt syndrome, scleritis,scleroderma, Sjogren's syndrome, sperm and testicular autoimmunity,stiff person syndrome, subacute bacterial endocarditis, Susac'ssyndrome, sympathetic ophthalmia, Takayasu's arteritis, temporalarteritis/giant cell arteritis, thrombocytopenic purpura, Tolosa-Huntsyndrome, transverse myelitis, Type 1 diabetes, ulcerative colitis,undifferentiated connective tissue disease, uveitis, vasculitis,vesiculobullous dermatosis, vitiligo, and Wegener's granulomatosis.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g. amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for. Unlessindicated otherwise, parts are parts by weight, molecular weight isweight average molecular weight, temperature is in degrees Celsius, andpressure is at or near atmospheric. Standard abbreviations may be used,e.g., bp, base pair(s); kb, kilobase(s); pl, picoliter(s); s or sec,second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); kb,kilobase(s); bp, base pair(s); nt, nucleotide(s); i.m.,intramuscular(ly); i.p., intraperitoneal(ly); i.n., intranasal(ly);i.v., intravenous(ly); s.c., subcutaneous(ly); M/ml, million units or10⁶ CFU/ml; M, million units/mouse or 10⁶ CFU/mouse; and the like.

Materials and Methods

The following materials and methods were used in the Examples describedbelow.

Materials

RPMI serum-free media was obtained from the Life Technologies(Burlington, Ontario, Canada). RPMI was supplemented with 5-10% fetalbovine serum, sodium pyruvate, Penicillin-streptomycin and2-mercapto-ethanol to make complete medium, which was used in variouscell cultures. ConA, PWM and LPS were obtained from Sigma ChemicalCompany. Cytokine kits and fluorescent-labeled anti-mouse antibodieswere purchased from eBioscience (San Diego, Calif.). Anti mouse FOXP3was purchased from biolegend (San Diego, Calif.). Aldra cream (5%imiquimod) was obtained University of Alberta Hospital pharmacy.Wild-type and lipopolysaccharide (LPS)-negative Caulobacter crescentus(LPS^(−ve) CC) were grown at room temperature (22-27° C.) in theincubator, and stored in saline at 4° C. or room temperature for varioustime periods. Wild-type Caulobacter vibroides (CV) was grown at roomtemperature (22-27° C.) in the incubator, and stored in saline at 4° C.or room temperature for various time periods.

Methods

Mice

6-8 Weeks old C57BL/6 or 9-11 week BALB/c, male or female mice werepurchased from Charles River Breeding Laboratories. All animalexperimental protocols used in this study were approved by theUniversity of Alberta Animal Care and Use Committee for Health Sciences,and conducted in accordance with the guidelines of the University ofAlberta, Edmonton, Canada

Treatments of Mice and Sample Collections

The mice were administered single or multiple times with Caulobactercrescentus (CC) in PBS using doses, schedule and routes as described indifferent examples and figure legends. CC used was prepared in twoformats: CC-1 (CC grown in liquid PYE medium and stored at roomtemperature in saline), and CC-2 (CC grown in liquid PYE medium andstored at 4° C. in refrigerator). CC-2 has also been referred to as CCin figures and description.

After euthanization of mice at specific times, blood, spleen, lungs,liver, lymph nodes, etc. were collected. Serum samples were used todetermine biochemical markers using commercial Vet test kits (IdexxLaboratories).

Isolation of Lymphocytes from Spleen

At specific times after immunization, the mice were euthanized to obtainsplenocytes. The spleens were pooled from 3-5 mice and ground to asingle cell suspension and filtered through a Falcon 100 μm nylon cellstrainer. After centrifugation, the cell pellet was resuspended in 2 mlof sterile distilled water and briefly vortexed. Immediately, 2×PBS wereadded and after a brief vortex the volume was made to 25 ml with 1×PBS.The tube was centrifuged and the cell pellet was resuspended in 10 ml ofcomplete RPMI. It was again filtered through a Falcon 100 μm nylon cellstrainer and centrifuged. The cell pellet was resuspended in 2 ml ofRPMI media. These lymphocytes were used for the experiment.

Mouse Cytokine ELISA

Cytokines and chemokines secreted in the supernatant of proliferatingco-cultures, or mouse serum samples were measured using sandwichenzyme-linked immunosorbent assay (ELISA) kits following themanufacturer's protocol (eBioscience, CA, USA) for the presence ofIL-10, GM-CSF, IL-17A, IFN-γ, TNF-α, IL-2, IL-6, IL-1βIL-22, MIP-1α,IL-8/KC. A dilution of 1:2 to 1:50 was used for the samples with thestandards ranging from 5 to 2,000 pg/ml. Finally the ELISA plates wereread and the concentrations were calculated with an automated ELISAplate reader (Fluostar Optima, BMG Labtech).

Evaluation of Antibody Responses

The levels of antibodies (IgG, IgG2a, IgE) in serum and lung washes weredetermined using enzyme-linked immunosorbent assays (ELISAs). Briefly,96-well nitrocellulose (Nunc) plates were coated with relevant antigen(such as OVA, MOG peptide) and incubated overnight at 4° C. The plateswere blocked with PBS containing normal mouse serum, followed byincubating with the experimental samples at different dilutions for 2hrs at room temperature. After washing the plates for 4 times, Antimouse Ig isotype antibodies conjugated with Alkaline phosphatase (AP)were added, followed by incubation for 2 hrs. After washing the plates,PNPP substrate was added and color development was read on FluostarELISA reader at 405 nm wavelength. All reagents for antibody detectionwere obtained from Southern Biotech (Birmingham, Ala.).

T Cell Proliferation Assay

Proliferative responses of splenic T cells were measured in triplicatecultures in 96-well flat-bottomed microtiter plates. A total of 4×10⁵spleen T cells from immunized mice and 4×10⁵ antigen-presenting cells(APCs) (spleen cells from control syngeneic mice irradiated with 18 Gy)were mixed with different concentrations (0.1, 1 and 10 μg/mL) of MOGpeptide were cultured in RPMI medium (with 10% fetal bovine serum (FBS))at 37° C. (5% CO₂) for 4 days.

The cells were pulsed with 0.5 μCi/well [³H]-thymidine (Amersham) for12-18 h and harvested on filter papers (Perkin Elmer). The levels of[³H]-thymidine incorporated into the DNA of proliferating cells werecounted in a Microbeta Trilux liquid scintillation counter (PerkinElmer). Proliferation is represented as the mean cpm±SE (standard error)of triplicate cultures.

Flow Cytometry Analysis of Surface Markers, Intracellular IL-10 andFoxp3

A total of 5×105 cells from immunized mice were taken for intracellularand extracellular staining with multicolor fluorescently labeled mAbs(concentrations according to manufacturer's instructions). The cellswere incubated with Fc mouse-serum (Sigma) to prevent non-specificbinding and washed with fluorescence-activated cell sorter (FACS)-buffer(2% fetal bovine serum in 1×phosphate-buffered saline (PBS). Afterincubation for 30 minutes with anti-mouse CD3e-FITC, CD4-PECy-5,CD25-PE-Cy7, CD8a-APC-Cy7, anti-PD-1-PerCP eFluore 710,anti-CD49b-Alexafluor-700 (for BALB/c and C57bl/6 mice) etc.(eBioscience) for extracellular markers at 4° C., the cells were washedtwice and fixed in fixative solution (1% paraformaldehyde inFACS-buffer) for 5 minutes. After washing twice, the cells wereincubated with cold permeabilization buffer (FACS-buffer+0.3% Saponin(Sigma)+5% normal human serum in PBS) for 5 minutes followed by additionof anti-mouse IL-10 (eBioscience) and anti-Foxp3-PE (biolegend) andfurther incubated for 30 minutes at 4° C. The cells were washed oncewith FACS-buffer containing 1% Saponin and fixed. They were read inFortessa and analyzed using FACS-DIVA software (Becton Dickinson,Mountain View, Calif.). Each marker was gated based on its respectiveisotype-matched control monoclonal antibodies.

Human PBMCs and DCs

Peripheral blood mononuclear cells (PBMCs) were obtained from normalhuman donors using Ficoll-Paque. To obtain dendritic cells (DCs),adherent PBMCs were cultured with recombinant GM-CSF and IL-4 for 5-6days in RPMI media, using procedures well established in the literature.Human PBMCs cultured with test materials for specified times asdescribed in individual examples were stained with antibodies againstCD34, CD45, CD11c and CD11b, labeled with various fluorophores obtainedcommercially (eBiosciences), using standard procedures.

Results

The following examples are intended to illustrate rather than limit thescope of the invention.

The immunomodulatory effects of CC were tested alone and with differentantigens in different models and indications via systemic and mucosalroutes as follows.

Example 1: Effect of Caulobacter crescentus (CC) on Modulation ofInflammatory Cytokines in Concanavalin a (ConA) Stimulated SplenocytesUpon Oral and Intranasal Administration of CC in Healthy Mice

C57/bl6 male mice were treated twice weekly orally or intranasally forfive times with CC at 500×106 CFU/mouse or PBS control. Mice wereeuthanized 8 days after the last treatment. The spleens isolated andex-vivo stimulated with T cell mitogen ConA at 1 μg/ml concentration for24 hrs. Supernatant were collected and cytokines (IFN-γ, TNF-α, IL-6,and IL-17A) were measured using ELISA (FIG. 1). These resultsdemonstrate that treatment with CC down-regulates production ofinflammatory cytokines in splenocytes upon ex-vivo stimulation with ConA and suggest the role of CC in suppressing inflammation mediated by Tcells from a variety of infections including viral, bacterial, fungal aswell as from environmental toxins, drug reactions and autoimmunedisorders.

Example 2: Effect of Caulobacter crescentus (CC) on Modulation ofInflammatory Cytokines from Pokeweed (PWM) Stimulated Splenocytes UponOral and Intranasal Administration of CC in Healthy Mice

C57/bl6 male mice were treated twice weekly orally or intranasally forfive times with CC at 500×106 CFU/mouse or PBS control. Mice wereeuthanized 8 days after the last treatment. The spleens of treated micewere isolated and ex-vivo stimulated with a B cell mitogen PWM at 0.1ug/ml concentration for 24 hrs. Supernatant were collected for cytokines(IFN-γ, TNF-α, IL-6 and IL-17A) measurements using ELISA (FIG. 2). Theseresults demonstrate that treatment with CC down-regulates production ofinflammatory cytokines in splenocytes upon ex-vivo stimulation with PWMand suggest the role of CC in reducing excessive production ofproinflammatory cytokines in infection and non-infection relatedsystemic inflammatory disorders.

Example 3: Effect of Caulobacter crescentus (CC) on Induction ofAnti-Inflammatory Cytokine IL-10 In Vivo Upon Oral and IntranasalAdministration

Groups of five C57/bl6 male mice were treated twice weekly orally orintranasally for five times with CC at 500×10⁶ CFU/mouse or PBS control.Mice were euthanized 8 days after the last treatment and spleens wereisolated. Splenocytes were cultured with media, ConA and PWM for 24 hr.Supernatant were collected for IL-10 measurement using ELISA (FIG.3A-FIG. 3C). The results obtained demonstrate that CC up-regulates IL-10production ex-vivo in splenocytes upon 24 hr culture with or withoutstimulants and suggest that CC can induce IL-10 production in vivo.These results also indicate systemic immunomodulatory andanti-inflammatory role of CC in reestablishing homeostatic balance ininflammatory conditions.

Overall, results described in FIG. 1, FIG. 2, and FIG. 3 suggest that CChas capacity to induce higher levels of the anti-inflammatory cytokineIL-10 and downregulate levels of inflammatory cytokines IFN-γ, TNF-α,IL-6 and IL-17A. Pro-inflammatory cytokines play a major role in thepathogenesis of many diseases and hence there is an interest indeveloping therapeutics to modulate their excessive production in arange of afflicted diseases.

Example 4: Effect of Caulobacter crescentus (CC) on IL-10 Productionfrom Ex Vivo LPS-Stimulated Splenocytes Isolated from CC Treated Mice

Female C57/bl6 mice were treated twice weekly orally via gavage for fourtimes with CC at 500×10⁶ CFU/mouse or PBS control. Mice were euthanized4 days after the last treatment and spleens were collected. Splenocyteswere stimulated with LPS at 1 μg/ml concentration for 24 and 72 hrs.Supernatants were collected and IL-10 was measured using ELISA (FIG. 4).The results obtained demonstrate that CC upregulates IL-10 productionex-vivo in splenocytes upon LPS stimulation compared to placebo group(PBS). These studies suggest the anti-inflammatory activity of CC innormalizing the dysregulated release of pro-inflammatory cytokines andprotection against inflammatory responses and/or autoimmune disorders.

Example 5: Effect of Caulobacter crescentus (CC) on Pro-InflammatoryCytokines in Gut-Associated Mesenteric Lymph Nodes Upon Oral Treatment

In order to determine the effect of CC on inflammatory cytokines levelsin intestinal lymphoid tissue, female C57/bl6 mice were treated twiceweekly orally for four times with CC at 500×10⁶ CFU/mouse or PBScontrol. Three days after the last treatment, mice were euthanized andlocal mesenteric lymph nodes were collected. Cytokines levels weredetermined in supernatant collected after 24 hr LPS stimulation. Oraladministration of CC led to decrease in the levels of IFN-γ, IL-6 andIL-17A compared to PBS group (FIG. 5). These results demonstrated thatCC can down-regulates production of Th1 and Th17 cytokines in localmesenteric lymph nodes. Thus, CC has ability to reduce TH1 and/or TH17mediated pro-inflammatory cytokine levels in a variety of inflammatorydisorders where immune dysregulation and inflammation is triggered byexternal or internal stimuli or microorgamisms such as Gram+ andGram-bacteria, viruses, fungi, parasites, LPS, toxins, autoantigens,glycosylphosphatidyl-inositol, tissue injury (trauma, burns) etc.

Example 6: Effect of Caulobacter crescentus (CC) in Systemic ImmuneModulation in Mice after Oral Treatment

Female C57/bl6 mice were treated orally twice weekly for four times withCC at 500×10⁶ CFU/mouse or PBS control. Mice were euthanized 4 daysafter the last treatment and spleens were collected. The percentage ofCD3+CD4+ and CD3+CD8+ cells expressing IL-10⁺ were analyzed by flowcytometry. Treatment with CC led to increased expression ofintracellular IL-10 on both CD4⁺ and CD8⁺ T cells in splenocytes (FIG.6). Taken together, the results shown in FIG. 4-FIG. 6 demonstrate thatCC has strong ability to normalize immune dysregulation both locally andsystemically in inflammatory and autoimmune disorders.

Example 7: Effect of Caulobacter crescentus (CC) on Modulation ofPro-Inflammatory Cytokine Production in Spleen after SubcutaneousTreatment

Groups of three C57/bl6 male mice were administered with CC (500×10⁶CFU/mouse) or PBS once weekly for four weeks by subcutaneous route. Micewere euthanized 28 days after the last treatment. Spleens were harvestedand splenocytes were cultured with ConA. Cytokines levels weredetermined in supernatant collected after 24 hr ConA stimulation. Theresults presented in FIG. 7 show that CC led to persistent modulation inthe production of IFN-γ, TNF-α and IL-6 cytokines (FIG. 7). Thus, CC canprovide long-lasting immunomodulatory effect even after cessation oftherapy.

Example 8: Modulation of Innate and Adaptive Immune Cells after OralTreatment with Caulobacter crescentus (CC)

Groups of three C57/bl6 male mice were treated orally with CC (500×10⁶CFU/mouse) or PBS once weekly for four weeks. Mice were euthanized 28days after the last treatment and splenocytes were harvested andanalyzed by flow cytometry. Oral treatment with CC led to increasedFOXP3 expression on CD8⁺, CD8⁺CD25⁺ and NKT (CD3⁺CD49b⁺) cells andincreased PD-1 expression on NK cells (FIG. 8). Thus CC induces variousregulatory lymphocytes to control inflammation. This data suggest thatCC can induce homeostasis by regulating the expression of differentregulatory molecules on innate and adaptive immune cells and therefore,CC can be used to control excessive inflammation in various diseases.PD-1 expression on NK cells has been shown to control inflammatoryresponses in mycobacterial infection. In addition, PD-1 on NK has alsoshown to protect from inflammation including viral, bacterial andautoimmune disorders.

NK and/or NKT cells have been shown in down regulating autoimmuneresponses in several diseases including multiple sclerosis, rheumatoidarthritis, systemic lupus erythematosus, Sjogren's syndrome, autoimmunethyroid disease, psoriasis, Behcet's disease, type I diabetes,neurodegenerative disease etc. Therefore, the present disclosurerepresents attractive biotherapeutic for the prevention and/or treatmentof a range of inflammatory, allergic and autoimmune disorders. Thus,overall the present disclosure represents a strategy to prevent and/ortreat systemic and local inflammation in infectious and non-infectioussettings through modulating the activity of adaptive and innate T and/orNK and/or NKT cells.

Example 9: Modulation of T Cells in Splenocytes after SubcutaneousTreatment with Caulobacter crescentus (CC)

Groups of three C57/bl6 male mice were treated subcutaneously with CC(500×10⁶ CFU/mouse) or PBS once weekly for four weeks. Mice wereeuthanized 28 days after the last treatment and splenocytes wereharvested and analyzed by flow cytometry. Subcutaneous treatment with CCled to increased FOXP3 expression on CD4⁺, CD4⁺CD25⁺, CD8⁺ and CD8⁺CD25⁺T cells (FIG. 9). Thus parenteral administration of CC can also providelong-lasting immunomodulatory effects. These results demonstrate theeffect of CC in inducing and expanding subsets of regulatory T cells. Tregulatory cells have been shown to suppress inflammatory activity in awide range of diseases including autoimmune encephalomyelitis, IBD,bacterial-induced colitis, type 1 diabetes, airway eosinophilicinflammation, graft vs. host disease, organ transplantation etc.

Example 10: Caulobacter crescentus (CC) Exhibits Positive Benefits inControlling Systemic Inflammation in LPS Challenged Model ofSepsis/Inflammation: Modulation of Cytokine Levels in Serum

Groups of 3 C57/bl6 male mice were challenged with LPS at 7 mg/Kg in 100μl PBS intraperitonially and treated orally with CC (500×10⁶ CFU/mouse)post 2 and 24 hr in vivo challenge with LPS. Healthy unchallenged andPBS fed mice were also included as controls. Mice were euthanized afterthe second treatment and blood samples were collected and analyzed forcytokines by ELISA. The LPS challenged mice had high levels ofinflammatory cytokines in sera. In contrast, CC treatment down-regulatedproduction of inflammatory cytokines IL-1β and IL-6 and up-regulated theproduction of anti-inflammatory cytokine IL-10 (FIG. 10). Thus, CCpromotes non-damaging immune responses. Induction of IL-1β and IL-6 hasbeen associated with a number of acute and chronic inflammatory andauto-immune diseases such as sepsis, MS, Alzheimers, Parkinsons, RA,gout, metabolic diseases (atherosclerosis, type II diabetes),hypertension, chronic obstructive pulmonary disease (COPD), asthma,psoriasis and allergy. Therefore, CC could help in managing andameliorating these inflammatory medical conditions.

Example 11: Caulobacter crescentus (CC) Exhibits Positive Benefits inControlling Local Inflammation in a Sepsis/Inflammation Model:Modulation of Cytokine Levels in Lungs and Liver

Groups of 3 C57/bl6 male mice were challenged with LPS at 7 mg/Kg in 100μl PBS intraperitonially and treated orally with CC (500×106 CFU/mouse)post 2 and 24 hr in vivo challenge with LPS. Healthy unchallenged andPBS fed mice were also included as controls. In these studies, CC usedwas prepared in two formats: CC-1 (CC grown in liquid PYE medium andstored at room temperature in saline), and CC-2 (CC grown in liquid PYEmedium and stored at 4° C. in refrigerator). Mice were euthanized after2nd treatment of CC-1 and CC-2, and lungs and liver were harvestedfollowed by determining cytokines in their homogenates. Both CC-1 andCC-2 down-regulated production of inflammatory cytokines TNF-α, IL-6,IL-1β and IL-17A in lungs and liver, compared to LPS challenged group(FIG. 11). These results demonstrate that CC can be utilized incontrolling inflammatory processes and normalizing tissue functionsincluding post-inflammmatory medical conditions related to viral andbacterial pathogens, tissue damage, cellular stress, metabolicperturbations etc. in various (intestine, lung, liver, brain, skin,heart etc.) organs. LPS mediated sepsis is a common cause of fatality incritical care, surgical and burn units. Therapeutic approaches directedtowards ameliorating LPS mediated fatal inflammatory cascade throughtargeting host immune components could have clinical and therapeuticadvantages.

Example 12: Caulobacter crescentus (CC) Exhibits Positive Effects onBiochemical Parameters of Liver Damage in LPS Challenged Mouse Model ofInflammation

Groups of 3 C57/bl6 male mice were challenged with LPS at 7 mg/Kg in 100μl PBS intraperitonially and treated orally with CC (500×10⁶ CFU/mouse)post 2 and 24 hr in vivo challenge with LPS. Healthy unchallenged andPBS fed mice were also included as controls. In these studies, CC usedwas prepared in two formats: CC-1 (CC grown in liquid PYE medium andstored at room temperature in saline), and CC-2 (CC grown in liquid PYEmedium and stored at 4° C. in refrigerator). Mice were euthanized after2nd treatment of CC-1 and CC-2, and blood samples were collected todetermine the effects on biochemical markers. Treatment with both CC-1and CC-2 normalized the serum levels of glucose (GLU), globulin (GLOB),alanine aminotransferases (ALT), total phosphates (TP) and totalbilirubin (TBIL) of LPS challenged mice to the levels of non-challengedand PBS fed mice (FIG. 12). These results demonstrate that CC could beused effectively in treating various inflammation mediated disordersincluding liver associated diseases.

Example 13: Caulobacter crescentus (CC) Prevents Liver Injury in LPSChallenged Mice Model

Groups of 3 C57/bl6 male mice were challenged with LPS at 7 mg/Kg in 100μl PBS intraperitonially and treated orally with CC (500×10⁶ CFU/mouse)post 2 and 24 hr in vivo challenge with LPS. Healthy unchallenged andPBS fed mice were also included as controls. In these studies, CC usedwas prepared in two formats: CC-1 (CC grown in liquid PYE medium andstored at room temperature in saline), and CC-2 (CC grown in liquid PYEmedium and stored at 4° C. in refrigerator). Mice were euthanized after2nd treatment of CC-1 and CC-2 and various organs were collected. Noinflammation or damage to lung, liver and other organs was observed withboth CC-1 and CC-2. H & E staining of liver sections were performed. Theresults shown in FIG. 13 demonstrate a protective effect of CCadministered by oral route in the prevention of LPS induced inflammationand liver damage, compared to LPS challenged mice. Massive destructionof liver has been associated with a number of medical conditions due toundesirable inflammatory activities as a result of infectious andnon-infectious pathogenesis. Therefore, CC has strong potential inpreventing and ameliorating liver damage from autoimmune hepatitis,alcohol related hepatic disorders, viral mediated hepatitis etc.

Example 14: Modulation of Cytokines in Imiquimod (IMQ)-InducedPsoriasis-Like Dermatitis Mouse Model Upon Oral Treatment withCaulobacter crescentus (CC)

10-11 Weeks old Balb/c female mice were administered a topical dose of6.25 mg of 5% IMQ cream (Aldra, Imiquimod) for 6 consecutive days onshaved back. Mice were treated orally with CC at 500×10⁶ CFU/mouse fromday 3 (5 days a week for two weeks). Mice were euthanized 3 days afterthe last treatment and spleens were harvested. Splenocytes were culturedex vivo with medium for 4 days and culture supernatants were examinedfor cytokines (FIG. 14). Treatment with CC led to a reduction in theproduction of IFN-γ, IL-6, IL-17A and IL-22 in Aldra induced psoriasismice compared to PBS treated psoriatic mice (FIG. 14). These studiessuggest that CC can correct cytokine dysregulation in inflammatory andautoimmune diseases and lead to beneficial therapeutic effects. Besidespsoriasis, infiltration of immune cells in the dermis and epidermis alsooccur in other chronic skin diseases such as atopic dermatitis, acneinversa, rosacea etc. Therefore, CC can be used in treating various skindisorders with an inflammatory component. Taken together, the resultspresented in FIG. 14 demonstrate that CC has systemic immunomodulatoryand anti-inflammatory activity to protect against pathogen or autoimmuneassociated inflammatory responses.

Example 15: Effect of Caulobacter Cescentus (CC) in ReducingPro-Inflammatory Cytokines and Chemokines in Colon Tissues ofDSS-Induced Inflammatory Bowel Disease (IBD) Afflicted Mice

Groups of 5 male C57bl/6 were given 3% dextran sulfate sodium (DSS) indrinking water for 7 consecutive days. Mice were treated with CC(500×10⁶ CFU/mouse) on the same day DSS was started. Water treated micewere used as controls. On day 10, mice were euthanized and colon tissueswere harvested and cultured for 24 h. Cytokines were determined inculture supernatants using ELISA. The results obtained indicate that CCreduced pro-inflammatory cytokines IFN-γ, TNF-α, IL-1β, and chemokinesIL-8/KC and MIP-1α(FIG. 15). Thus, oral treatment of CC attenuatedaberrant levels of inflammatory cytokines and chemokines locally incolon in the murine DSS-induced colitis model for IBD, which account foractivation of cascades leading to epithelial permeability, apoptosis,ulceration, diarrhea etc. in IBD.

Example 16: Effect of Caulobacter crescentus (CC) in SuppressingAuto-Antigen Specific T Cell and Antibody Responses in ExperimentalAutoimmune Encephalomyelitis (EAE) Model

To determine the effect of CC in reducing auto-antigen specific immuneresponses, groups of five C57Bl6 female mice were immunized with 200 μmMOG₃₅₋₅₅ peptide emulsified in CFA in 100 μl saline subcutaneously.Additionally, mice received 400 ng of pertussis toxin in 200 μl salineintraperitoneally at day 0 and 3. Starting from day 3 of immunization,mice were treated orally with CC (500×10⁶ cfu/mouse) continuously every3^(rd) day till the end of the experiment. PBS treated challenged micewere used as controls. Mice were euthanized 30 days after immunizationand spleen and blood samples were collected. T cell responses wereexamined in splenocytes against MOG peptide (10, 1 and 0.1 μg/ml).Anti-MOG antibody (IgG and IgG2a) titers were analyzed by ELISA in serumsamples. Interestingly, CC reduced autoantigen-MOG-specific T cellresponses compared to PBS control group (FIG. 16). CC also reduced theautoantigen-MOG-specific IgG and IgG2a antibody titers compared to PBScontrol group (FIG. 16). These results suggest that CC can reduceautoantigen specific T and B cell responses. EAE model is a commonlyused model for the human inflammatory demyelinating disease, multiplesclerosis (MS). MS is an inflammatory disease of the central nervoussystem (CNS). EAE is a complex condition where interaction betweenimmunopathological and neuropathological mechanisms leads topathological features of MS, viz., inflammation, demyelination, axonalloss and gliosis. Therefore, CC can be effective in treating autoimmuneand neurological disorders, autoimmune blood diseases, autoimmunehemolytic anemia etc.

Example 17: Effect of Caulobacter crescentus (CC) in Reducing Allergen(OVA)-Specific Antibody and Cytokine Responses in a Mouse Model ofOvalbumin-Induced Airway Inflammation Model

To determine the effect of CC in airway/lung inflammatory diseases,groups of 5 Balb/c male mice were sensitized on day 1 and day 6 with 10μg ovalbumin and 2 mg Al(OH)₃ in 400 μl saline intraperitoneally. Micewere challenged intranasally (15 μl/nostril) with 50 μg ovalbumin ondays 12 and 14. From day 7 onwards of sensitization, mice were treatedorally with CC (500×10⁶ CFU/mouse) every third day up to day 17. Inseparate groups, mice were treated with dexamethasone (DEX, 2 mg/Kg,i.p.) alone once on day 13 or dexamethasone and CC. Control micereceived saline at the same schedule as CC. Mice were euthanized nextday of the last treatment, blood samples, lung washes and spleen werecollected. A single cell-suspension (2×10⁶ cells/ml) of splenocytes wascultured with 50 μg OVA for 96 h and cytokines (IL-4 and IL-6) levelswere analyzed in culture supernatants by ELISA. Oral treatment with CCreduced the production of OVA specific IgEs in serum and lung washes inmice sensitized with OVA (FIG. 17). It was found that levels of IL-4 andIL-6 were also reduced in spleen by CC treatment (FIG. 18). Combining CCtreatment with dexamethasone led to further reduction in IL-4 and IL-6levels in spleen compared to dexamethasone alone treatment (FIG. 19).These data suggest that treatment with CC prevents OVA-induced allergicairway inflammation in mice. Allergy is a complex disease where multipleimmune cells and inflammatory mediators contribute to the initiation andmanifestation of allergic diseases. Allergies are most frequently theresults of IgE-mediated hypersensitivity reactions. Increased productionof IL-4 and IL-6 has been correlated with the allergic diseases. Thepresence of IgE in serum is a hallmark of allergic diseases driven byIL-4 mediated Ig class switching in B cells. IgE dependent allergicreactions may affect one or more target organs and allergic diseasessuch as rhinitis, sinusitis, conjunctivitis, asthma, dermatitis, andfood allergies etc.

Example 18: Effect of Caulobacter crescentus (CC) in Treating MetabolicDisorders and Reducing Systemic Inflammation in High-Fat Diet InducedObesity Model

Groups of five C57bl/6 mice were fed high-fat diet for 4-6 months andtreated with CC (500×10⁶ cfu/mouse) thrice weekly for 4-6 months. PBStreated high-fat diet fed mice were used as controls. Glucose tolerancetest was performed prior to terminating the experiment at ˜180 days.Blood glucose was measured using OneTouch monitoring system. Mice wereeuthanized and blood, liver and spleen were collected. Serum sampleswere used to determine biochemical parameters by Vet test. Liverhomogenates were prepared and pro-inflammatory cytokines (IFN-γ, TNF-α,IL-6, IL-1β) were determined by ELISA. Splenocytes (2×10⁶ cells/ml) werecultured with LPS (1 mg/ml) for 24 h and cytokines (IFN-γ, TNF-α, IL-6)were analyzed in culture supernatants by ELISA. Oral treatment with CCsignificantly reduced pro-inflammatory cytokines in liver (FIG. 20) andspleen (FIG. 21) compared to high-fat diet control mice, suggesting thatCC can effectively reduce chronic and systemic inflammation. Oraltreatment with CC also provided positive benefits in biochemicalparameters associated with metabolic diseases in high-fat diet inducedobesity model (FIG. 22). Further mice on CC treatment demonstratedimproved glucose tolerance compared to control group (FIG. 23). It hasbeen well established that obesity is associated with low-gradeinflammation contributing to a number of metabolic disorders includingtype 2 diabetes mellitus, cardiovascular diseases, hypertension,hypercholesterolemia, hypertriglyceridemia, non-alcoholic fatty liverdisease, hair loss etc. In obese state, production of a number ofinflammatory cytokines and chemokines (e.g., IFN-γ, TNF-α, IL-6, IL-1βetc.) is dysregulated. Obesity is also associated with insulinresistance leading to glucose intolerance, and non-alcoholic fatty liverdisease. It has been shown that inhibition of the activity of TNF-α alsosignificantly inhibits the development of steatohepatitis in alcohol-fedanimals. High cholesterol is associated with elevated risks of coronaryheart disease (atherosclerosis), stroke, peripheral vascular diseases,diabetes, and high blood pressure. Elevated levels of triglycerides areassociated with atherosclerosis and also increase the risk of acutepancreatitis. A high uric acid level occurs when kidneys do noteliminate uric acid efficiently. Other factors that may cause high uricacid include obesity, hypothyroidism etc. Build up of uric acid can leadto inflammation and intense pain of a gout attack. An elevated level ofcreatine kinase is seen in heart attacks, all types of musculardystrophy, endocrine disorders, neuromuscular diseases etc.Pro-inflammatory cytokines such as TNF-α, IL-6, IFN-γ, IL-1β are knownto be involved in obesity, obesity-related pathologies such as type 2diabetes, insulin-resistance, non-alcoholic fatty liver disease,alcohol-induced liver disease, vascular dysfunction relatedneurodegenerative processes such as Alzheimer disease, neuropsychiatricdisorders such as depression, schizophrenia etc. Mitochondrial reactiveoxygen species (ROS) drive pro-inflammatory cytokine (TNF-α, IL-6,IFN-γ, IL-1β) production in chronic human diseases. These studies showthat treatment with CC reduces serum biochemical markers associated withmetabolic disorders, improves glucose tolerance and reducespro-inflammatory cytokines in liver in obese mice on high-fat diet.

Example 19: Effect of Caulobacter crescentus (CC) in Reducing Hepato-and Nephro-Toxicities Associated with Anticancer Drug Cyclophosphamidein EL-4 Lymphoma-Bearing Mice

To determine the effect of CC in reducing toxicities associated with atherapeutic treatment, groups of five C57Bl6 mice were challenged with0.25×10⁶ EL-4 cells/mouse in 100 μl saline subcutaneously in the lowerleft flank. Starting from day 5 post challenge with EL-4 cells, CC(50×10⁶ cfu/mouse) was administered once weekly orally. On daysseventeen and twenty-one, mice were treated with cyclophosphamide at 150mg/Kg intraperitoneally. Healthy unchallenged and cyclophosphamide alonetreated challenged mice were used as controls. Mice were humanelyeuthanized 30 days after EL-4 challenge and blood samples werecollected. Treatment with CC along with cyclophosphamide normalized theserum levels of phosphate (PHOS) and total bilirubin (TBIL) in EL-4tumor bearing mice to the levels of non-challenged normal mice (FIG.24). These results demonstrate that CC could be used effectively inreducing toxicities of anticancer agents or a therapeutic treatment.

Example 20: Effect of Caulobacter crescentus (CC) in Reducing OrganToxicities Associated with Anticancer Drug Cisplatin in B16 MetastaticCancer Model

To determine the effect of CC in reducing toxicity associated withcancer chemotherapy, groups of five C57Bl6 mice were challenged with0.4×10⁶ B16 cells/mouse in 100 μl saline intraperitoneally. Mice weretreated with cisplatin (4 mg/kg) intraperitoneally at days 7 and 10 postB16 challenge and CC (50×10⁶ cfu/mouse) was administered orally onceweekly. Healthy unchallenged and cisplatin alone treated challenged micewere used as controls. Mice were euthanized 30 days after B16 challengeand blood samples were collected to determine the effects on serumbiochemical markers. Treatment with CC along with cisplatin normalizedthe serum levels of phosphate (PHOS), urea, total bilirubin (TBIL) andaspartate aminotransferase (AST) in B16 metastatic cancer bearing miceto the levels of non-challenged normal mice (FIG. 25). These resultsdemonstrate that CC could be used effectively in reducing toxicities ofanticancer agents or a therapeutic treatment.

Example 21: Effect of Caulobacter crescentus (CC) in Reducing ElevatedLevels of Biochemical Parameters of Hepatotoxicity Associated withCheckpoint Inhibitor Antibody (Anti-PD-1) in Mouse Model of B16 Melanoma

To determine the effect of CC in reducing toxicities associated withtherapeutic antibodies, groups of five C57Bl6 mice were challenged with0.4×10⁶ B16 cells/mouse in 100 μl saline subcutaneously in the lowerleft flank. Starting from day 3 post challenge with B16 melanoma cancercells, CC (50×10⁶ cfu/mouse) was administered orally once weekly. Twodays after first treatment with CC, anti-PD-1 antibody (200 ug/mouse)was administered intraperitoneally and continued every 3-4 days. Healthyunchallenged and anti-PD-1 antibody alone treated challenged mice wereused as controls. Mice were euthanized 25 days after tumor challenge andblood samples were collected to determine the effects on biochemicalmarkers. Treatment with CC along with anti-PD-1 monoclonal antibodynormalized the serum levels of phosphate alanine aminotransferases (ALT)and aspartate aminotransferase (AST) in B16 tumor bearing mice to thelevels of non-challenged normal mice (FIG. 26). These resultsdemonstrate that CC could be used effectively in reducing toxicities oftherapeutic and checkpoint inhibitor antibodies or a therapeutictreatment.

Example 22: LPS Negative Caulobacter crescentus (LPS' CC) Protects Micefrom Liver Damage in LPS Challenged Model of Sepsis/Inflammation:Modulation of Cytokine Levels in Liver

To determine the role of lipopolysaccharide negative (LPS^(−ve)) CC inimmune modulation, C57/bl6 female mice were challenged with LPS at 25mg/Kg in 200 μl saline intraperitonially and treated orally with LPS' CC(500×10⁶ CFU/mouse) post 2 and 20 h in vivo challenge with LPS. Healthyunchallenged and PBS fed mice were also included as controls. Mice wereeuthanized after 24 h of the second treatment, and liver was harvestedfollowed by determining cytokines in their homogenates. LPS challengedmice had high levels of inflammatory cytokines and chemokines in liverhomogenate. In contrast, LPS' CC treatment down-regulated production ofinflammatory cytokines IFN-γ, TNF-α, IL-1β, IL-6, IL-8/KC, and chemokineMIP-1α and up-regulated the production of anti-inflammatory cytokineIL-10 (FIG. 27). These results demonstrate that LPS^(−ve) CC alsoexhibits positive effect in controlling inflammatory processes, similarto those obtained using wild-type CC. Therefore, LPS negative CC can beutilized in controlling inflammatory processes and normalizing tissuefunctions including post-inflammatory medical conditions.

Example 23: Effect of Caulobacter vibroides (CV) in ReducingPro-Inflammatory Cytokines Induced by a Probiotic or Pathogenic Bacteriain Human PBMCs

Human PBMCs (4×10⁶/well) were stimulated with inactivated Lactobacillusrhamnosus (LB) or Lysteria monocytogenes (LM) (50×10⁶ CFU/ml) for 6 h.After that CV (250×10⁶ CFU/ml) was added and plates were incubated foradditional 24 or 96 h. Culture supernatants were collected and assayedfor IFN-γ and TNF-α by ELISA. Treatment with CV reduced the levels ofIFN-γ and TNF-α induced by LB and LM (FIG. 28). These results suggestthat other species of Caulobacter can reduce the inflammation related tobacterial or viral pathogens. These results also suggest thatCaulobacter species can be used to treat undesirable inflammatoryresponses induced by microbiome related and unrelated microbes. Further,they can be used to modulate immunomodulatory activity of a microbiometherapeutic.

Example 24: Effect of Caulobacter Cescentus (CC) in Ex Vivo Manipulationof Human Myeloid Dendritic Cells

Human myeloid dendritic cells differentiated from peripheral bloodmonocytes (2×10⁶/ml) were cultured for 24 h in the absence and presenceof CC (50 or 10 CFU/ml). Culture supernatants were collected andexamined for IL-10 levels by ELISA. The results obtained suggest that CCcan generate a population of modulated DCs producing IL-10 (FIG. 29).Dendritic cells (DCs) are key regulators of adaptive immunity with thepotential to induce T cell suppression and tolerance. IL-10 produced byDCs plays a role in generation, expansion and maintenance of regulatoryT cells. These results suggest that CC can be used for ex vivomanipulation of DCs for DC-based immunotherapeutic strategies.

Example 25: Effect of Caulobacter Cescentus (CC) onDifferentiation/Expansion of Pluripotent Stem Cells from Human PBMCsinto Myeloid Cells

Human PBMCs (4×10⁶/well) were cultured with CC (500×10⁶, 50×10⁶, 10×10⁶and 1×10⁶ CFU/ml) and saline for 10 days. PBMCs were stained for surfacemarkers CD34, CD45, CD11c and CD11b. Cells were gated forCD34+CD45-pluripotent stem cells, which were further gated for CD11c+and CD11b+ DCs and macrophages, and data were analyzed by flowcytometry. The results obtained suggest that CC can differentiate stemcells into myeloid cells (FIG. 30). Myeloid cells originate frommultipotent hematopoietic stem cells. These cells play a critical rolein innate and adaptive immunity, inflammatory reaction, restoration ofimmune homeostasis, bone remodelling etc. Thus, CC can lead todifferentiation and/or expansion of myeloid cells from stem cells foruse in patient specific immunotherapy.

What is claimed is:
 1. A method of treating an inflammatory disease,disorder or condition in an individual, the method comprisingadministering to the individual an effective amount of animmunomodulatory composition, the immunomodulatory compositioncomprising: a) live Caulobacter spp.; and b) a pharmaceuticallyacceptable excipient; wherein the composition is administered via anoral, nasal, or topical route of administration; wherein the Caulobacterspp. is selected from the group consisting of Caulobacter crescentus,lipopolysaccharide-negative Caulobacter crescentus, S-layer-negativeCaulobacter crescentus, Caulobacter vibroides, Caulobacter subvibriodes,Caulobacter henricii, Caulobacter fusiformis, and Caulobacterintermedius.
 2. The method of claim 1, further comprising administeringto the individual an anti-microbial agent, an anti-inflammatory agent,an anti-proliferative agent, a cytotoxic agent, an immunosuppressiveagent, an immunomodulatory agent, or an antibody.
 3. The method of claim1, further comprising administering to the individual a therapeuticmicrobe.
 4. The method of claim 1, wherein the individual is a human, anon-human mammal, or a non-mammalian animal.
 5. The method of claim 1,where the disorder or condition is selected from the group consisting ofrheumatoid arthritis, diabetes mellitus, multiple sclerosis, systemiclupus erythematosus, graft vs. host disease, fatty liver disease, liverfibrosis, steatohepatitis, pancreatitis, celiac disease, osteoporosis,alopecia areata, thyroiditis, obesity, atherosclerosis, asthma,pulmonary fibrosis, chronic obstructive pulmonary disease, inflammatorybowel disease, mucositis, psoriasis, atopic dermatitis, eczema, sepsis,and nephritis.